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Gromeier, Matthias

Overview:

Neuro-Oncology
Protein Synthesis Regulation
Signal Transduction
Growth & Proliferation Control in Cancer
Oncolytic Viruses
Viral Neuropathogenesis
Immunization Vectors

Positions:

Professor of Neurosurgery

Neurosurgery
School of Medicine

Professor in Molecular Genetics and Microbiology

Molecular Genetics and Microbiology
School of Medicine

Professor in Medicine

Medicine, Infectious Diseases
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

M.D. 1992

M.D. — University of Hamburg (Germany)

Postdoctoral Fellow, Molecular Genetics & Microbiology

State University of New York at Stony Brook

Postdoctoral Associate, Molecular Genetics & Microbiology

State University of New York at Stony Brook

News:

Grants:

Oncolytic Polovirus, Immunotoxin, and Checkpoint Inhibitor Therapy of Gliomas

Administered By
Pathology
AwardedBy
National Institutes of Health
Role
Co Investigator
Start Date
August 01, 2015
End Date
July 31, 2022

Regional Oncolytic Poliovirus Immunotherapy for Breast Cancer

Administered By
Surgery, Surgical Sciences
AwardedBy
Department of Defense
Role
Co Investigator
Start Date
August 01, 2016
End Date
July 31, 2021

Viral Oncology Training Grant

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Participating Faculty Member
Start Date
July 01, 1980
End Date
June 30, 2019

Combining PVSRIPO Immunotherapy with Lomustine for the Treatment of Recurrent GBM

Administered By
Neurosurgery
AwardedBy
National Brain Tumor Society
Role
Principal Investigator
Start Date
July 01, 2017
End Date
July 31, 2018

Cancer Immunotherapy Through Intratumoral Activation of Recall Responses

Administered By
Neurosurgery
AwardedBy
National Cancer Center
Role
Principal Investigator
Start Date
June 01, 2017
End Date
May 31, 2018

Targeting Translation Control in Malignant Glioma

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 17, 2007
End Date
May 31, 2018

Oncolytic Immunotherapy of Glioblastoma with Recombinant Poliovirus

Administered By
Neurosurgery
AwardedBy
Blast Glioblastoma
Role
Principal Investigator
Start Date
April 01, 2015
End Date
April 01, 2018

Phase-1 clinical trial of PVSRIPO oncolytic immunotherapy in pediatric HGG

Administered By
Neurosurgery
AwardedBy
Solving Kids' Cancer
Role
PD/PI
Start Date
April 01, 2016
End Date
March 31, 2018

Oncolytic PVSRIPO Expressing Tumor Antigens as a Cancer Vaccine

Administered By
Neurosurgery
AwardedBy
Oligo Nation
Role
Principal Investigator
Start Date
September 15, 2016
End Date
September 05, 2017

Research Training In Neuro-Oncology

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
July 01, 1998
End Date
August 31, 2016

Phase II/III Manufacture of the Oncolytic Poliovirus Chimera, PVSRIPO

Administered By
Neurosurgery
AwardedBy
North Carolina Biotechnology Center
Role
Principal Investigator
Start Date
August 03, 2015
End Date
August 02, 2016

Cancer Biology Training Grant

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Cancer Institute
Role
Mentor
Start Date
July 01, 1993
End Date
March 31, 2016

Oncolytic Poliovirus Immunotherapy of Malignant Glioma

Administered By
Pathology
AwardedBy
The Slomo and Cindy Silvian Foundation, Inc
Role
Co Investigator
Start Date
December 12, 2014
End Date
December 11, 2015

Oncolytic Virotherapy of Meningeal Cancer

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
July 01, 2009
End Date
May 31, 2015

Instrumentation for Quantitative Phosphoproteomics and Acetylomics

Administered By
Duke Center for Genomic and Computational Biology
AwardedBy
National Institutes of Health
Role
Major User
Start Date
May 15, 2014
End Date
May 14, 2015

Oncolytic poliovirus therapy of malignant glioma

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
Southeastern Brain Tumor Foundation
Role
Principal Investigator
Start Date
May 01, 2014
End Date
April 30, 2015

Enterovirus Vectors with Respiratory Tropism for Cancer Immunotherapy

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
September 15, 2009
End Date
August 31, 2012

Transgenic Mouse Model for the Common Cold

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
January 01, 2008
End Date
December 31, 2010

Research Training In Neuro-Oncology

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
September 15, 2005
End Date
August 31, 2010

Cis and Trans-acting Factors in HCV Gene Expression

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
December 01, 2005
End Date
November 30, 2008

Cell Type-Specific Viral Translation in the CNS

Administered By
Neurosurgery, Neuro-Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 21, 2002
End Date
July 31, 2007

Molecular Targeting of Breast Cancer

Administered By
Molecular Genetics and Microbiology
AwardedBy
United States Army Medical Research and Materiel Command
Role
Principal Investigator
Start Date
August 06, 2004
End Date
September 05, 2005

Cell Type-Specific Viral Translation in the CNS

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 01, 2003
End Date
July 31, 2005

Genetically stable Picornavirus HIV Immunization vectors

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
March 01, 2003
End Date
February 28, 2005

Production of non-pathogenic oncolytic poliovirus chimeras for the treatment of brain tumors (Project #417)

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
November 01, 2001
End Date
April 01, 2002

Production of non-pathogenic oncolytic poliovirus chimeras for the treatment of brain tumors

Administered By
Molecular Genetics and Microbiology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
November 07, 2000
End Date
May 30, 2001
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Publications:

Cancer immunotherapy with recombinant poliovirus induces IFN-dominant activation of dendritic cells and tumor antigen-specific CTLs.

Tumors thrive in an immunosuppressive microenvironment that impedes antitumor innate and adaptive immune responses. Thus, approaches that can overcome immunosuppression and engage antitumor immunity are needed. This study defines the adjuvant and cancer immunotherapy potential of the recombinant poliovirus/rhinovirus chimera PVSRIPO. PVSRIPO is currently in clinical trials against recurrent World Health Organization grade IV malignant glioma, a notoriously treatment-refractory cancer. Cytopathogenic infection of neoplastic cells releases the proteome and exposes pathogen- and damage-associated molecular patterns. At the same time, sublethal infection of antigen-presenting cells, such as dendritic cells and macrophages, yields potent, sustained type I interferon-dominant activation in an immunosuppressed microenvironment and promotes the development of tumor antigen-specific T cell responses in vitro and antitumor immunity in vivo. PVSRIPO's immune adjuvancy stimulates canonical innate anti-pathogen inflammatory responses within the tumor microenvironment that culminate in dendritic cell and T cell infiltration. Our findings provide mechanistic evidence that PVSRIPO functions as a potent intratumor immune adjuvant that generates tumor antigen-specific cytotoxic T lymphocyte responses.

Authors
Brown, MC; Holl, EK; Boczkowski, D; Dobrikova, E; Mosaheb, M; Chandramohan, V; Bigner, DD; Gromeier, M; Nair, SK
MLA Citation
Brown, MC, Holl, EK, Boczkowski, D, Dobrikova, E, Mosaheb, M, Chandramohan, V, Bigner, DD, Gromeier, M, and Nair, SK. "Cancer immunotherapy with recombinant poliovirus induces IFN-dominant activation of dendritic cells and tumor antigen-specific CTLs." Science translational medicine 9.408 (September 2017).
PMID
28931654
Source
epmc
Published In
Science Translational Medicine
Volume
9
Issue
408
Publish Date
2017
DOI
10.1126/scitranslmed.aan4220

MNK Controls mTORC1:Substrate Association through Regulation of TELO2 Binding with mTORC1.

The mechanistic target of rapamycin (mTOR) integrates numerous stimuli and coordinates the adaptive response of many cellular processes. To accomplish this, mTOR associates with distinct co-factors that determine its signaling output. While many of these co-factors are known, in many cases their function and regulation remain opaque. The MAPK-interacting kinase (MNK) contributes to rapamycin resistance in cancer cells. Here, we demonstrate that MNK sustains mTORC1 activity following rapamycin treatment and contributes to mTORC1 signaling following T cell activation and growth stimuli in cancer cells. We determine that MNK engages with mTORC1, promotes mTORC1 association with the phosphatidyl inositol 3' kinase-related kinase (PIKK) stabilizer, TELO2, and facilitates mTORC1:substrate binding. Moreover, our data suggest that DEPTOR, the endogenous inhibitor of mTOR, opposes mTORC1:substrate association by preventing TELO2:mTORC1 binding. Thus, MNK orchestrates counterbalancing forces that regulate mTORC1 enzymatic activity.

Authors
Brown, MC; Gromeier, M
MLA Citation
Brown, MC, and Gromeier, M. "MNK Controls mTORC1:Substrate Association through Regulation of TELO2 Binding with mTORC1." Cell reports 18.6 (February 2017): 1444-1457.
PMID
28178522
Source
epmc
Published In
Cell Reports
Volume
18
Issue
6
Publish Date
2017
Start Page
1444
End Page
1457
DOI
10.1016/j.celrep.2017.01.023

Recombinant oncolytic poliovirus, PVSRIPO, has potent cytotoxic and innate inflammatory effects, mediating therapy in human breast and prostate cancer xenograft models.

Intratumoral inoculation of viruses with tumor-selective cytotoxicity may induce cancer cell death and, thereby, shrink neoplastic lesions. It is unlikely, however, that viral tumor cell killing alone could produce meaningful, durable clinical responses, as clinically suitable 'oncolytic' viruses are severely attenuated and their spread and propagation are opposed by host immunity. Thus, a more propitious event in this context is the innate antiviral response to intratumoral virus administration, in particular for recruiting durable adaptive immune effector responses. It may represent a double-edged sword, as innate immune activation may eliminate infected tumor cells early, intercept viral spread and block any meaningful therapeutic response. The innate response to viral infection of tumors may be very different from that in non-malignant target tissues, owing to the unusual composition/tissue properties of tumor stroma. In this work, we report investigations of the innate immune response to the oncolytic poliovirus recombinant, PVSRIPO, in two mouse xenotransplantation models for breast and prostate cancer. Our observations indicate short-term virus persistence in infected tumors and virus recovery indicative of modest intratumoral propagation and persistence. Yet, a powerful innate inflammatory response coincided with chemokine induction and myeloid cell infiltration into tumors that was, interestingly, dominated by neutrophils. The combined effect of PVSRIPO tumor infection and the innate response it elicits was significant tumor regression in both models.

Authors
Holl, EK; Brown, MC; Boczkowski, D; McNamara, MA; George, DJ; Bigner, DD; Gromeier, M; Nair, SK
MLA Citation
Holl, EK, Brown, MC, Boczkowski, D, McNamara, MA, George, DJ, Bigner, DD, Gromeier, M, and Nair, SK. "Recombinant oncolytic poliovirus, PVSRIPO, has potent cytotoxic and innate inflammatory effects, mediating therapy in human breast and prostate cancer xenograft models." Oncotarget 7.48 (November 2016): 79828-79841.
PMID
27806313
Source
epmc
Published In
Oncotarget
Volume
7
Issue
48
Publish Date
2016
Start Page
79828
End Page
79841
DOI
10.18632/oncotarget.12975

Cytotoxic and immunogenic mechanisms of recombinant oncolytic poliovirus.

An oncolytic virus (OV) based on poliovirus (PV), the highly attenuated polio/rhinovirus recombinant PVSRIPO, may deliver targeted inflammatory cancer cell killing; a principle that is showing promise in clinical trials for recurrent glioblastoma (GBM). The two decisive factors in PVSRIPO anti-tumor efficacy are selective cytotoxicity and its in situ immunogenic imprint. While our work is focused on what constitutes PVSRIPO cancer cytotoxicity, we are also studying how this engenders host immune responses that are vital to tumor regression. We hypothesize that PVSRIPO cytotoxicity and immunogenicity are inextricably linked in essential, complimentary roles that define the anti-neoplastic response. Herein we delineate mechanisms we unraveled to decipher the basis for PVSRIPO cytotoxicity and its immunotherapeutic potential.

Authors
Brown, MC; Gromeier, M
MLA Citation
Brown, MC, and Gromeier, M. "Cytotoxic and immunogenic mechanisms of recombinant oncolytic poliovirus." Current opinion in virology 13 (August 2015): 81-85. (Review)
PMID
26083317
Source
epmc
Published In
Current Opinion in Virology
Volume
13
Publish Date
2015
Start Page
81
End Page
85
DOI
10.1016/j.coviro.2015.05.007

Oncolytic immunotherapy through tumor-specific translation and cytotoxicity of poliovirus.

Achieving tumor-specific, robust, and durable effector cytotoxic immune responses is key to successful immunotherapy. This has been accomplished with adoptive cell transfer of ex vivo-expanded autologous tumor-infiltrating or engineered T cells, or with immune checkpoint inhibitors, enhancing inherent T cell reactivity. A natural ability to recruit effector responses makes tumor-targeting ('oncolytic') viruses attractive as immunotherapy vehicles. However, most viruses actively block inflammatory and immunogenic events; or, host innate immune responses may prevent immune initiating events in the first place. Moreover, the mechanisms of how virus infection can produce effector responses against host (tumor) neo-antigens are unclear. We are pioneering oncolytic immunotherapy based on poliovirus, which has no specific mechanism to interfere with host immune activation, exhibits lytic cytotoxicity in the presence of an antiviral interferon response and pre-existing immunity, and engages a powerful innate immune sensor implicated in recruiting cytotoxic T cell responses. Central to this approach is a unique confluence of factors that drive tumor-specific viral translation and cytotoxicity.

Authors
Brown, MC; Gromeier, M
MLA Citation
Brown, MC, and Gromeier, M. "Oncolytic immunotherapy through tumor-specific translation and cytotoxicity of poliovirus." Discovery medicine 19.106 (May 2015): 359-365. (Review)
PMID
26105699
Source
epmc
Published In
Discovery medicine
Volume
19
Issue
106
Publish Date
2015
Start Page
359
End Page
365

Oncolytic polio virotherapy of cancer.

Recently, the century-old idea of targeting cancer with viruses (oncolytic viruses) has come of age, and promise has been documented in early stage and several late-stage clinical trials in a variety of cancers. Although originally prized for their direct tumor cytotoxicity (oncolytic virotherapy), recently, the proinflammatory and immunogenic effects of viral tumor infection (oncolytic immunotherapy) have come into focus. Indeed, a capacity for eliciting broad, sustained antineoplastic effects stemming from combined direct viral cytotoxicity, innate antiviral activation, stromal proinflammatory stimulation, and recruitment of adaptive immune effector responses is the greatest asset of oncolytic viruses. However, it also is the source for enormous mechanistic complexity that must be considered for successful clinical translation. Because of fundamentally different relationships with their hosts (malignant or not), diverse replication strategies, and distinct modes of tumor cytotoxicity/killing, oncolytic viruses should not be referred to collectively. These agents must be evaluated based on their individual merits. In this review, the authors highlight key mechanistic principles of cancer treatment with the polio:rhinovirus chimera PVSRIPO and their implications for oncolytic immunotherapy in the clinic.

Authors
Brown, MC; Dobrikova, EY; Dobrikov, MI; Walton, RW; Gemberling, SL; Nair, SK; Desjardins, A; Sampson, JH; Friedman, HS; Friedman, AH; Tyler, DS; Bigner, DD; Gromeier, M
MLA Citation
Brown, MC, Dobrikova, EY, Dobrikov, MI, Walton, RW, Gemberling, SL, Nair, SK, Desjardins, A, Sampson, JH, Friedman, HS, Friedman, AH, Tyler, DS, Bigner, DD, and Gromeier, M. "Oncolytic polio virotherapy of cancer." Cancer 120.21 (November 2014): 3277-3286. (Review)
PMID
24939611
Source
epmc
Published In
Cancer
Volume
120
Issue
21
Publish Date
2014
Start Page
3277
End Page
3286
DOI
10.1002/cncr.28862

Induction of viral, 7-methyl-guanosine cap-independent translation and oncolysis by mitogen-activated protein kinase-interacting kinase-mediated effects on the serine/arginine-rich protein kinase.

Protein synthesis, the most energy-consuming process in cells, responds to changing physiologic priorities, e.g., upon mitogen- or stress-induced adaptations signaled through the mitogen-activated protein kinases (MAPKs). The prevailing status of protein synthesis machinery is a viral pathogenesis factor, particularly for plus-strand RNA viruses, where immediate translation of incoming viral RNAs shapes host-virus interactions. In this study, we unraveled signaling pathways centered on the ERK1/2 and p38α MAPK-interacting kinases MNK1/2 and their role in controlling 7-methyl-guanosine (m(7)G) "cap"-independent translation at enterovirus type 1 internal ribosomal entry sites (IRESs). Activation of Raf-MEK-ERK1/2 signals induced viral IRES-mediated translation in a manner dependent on MNK1/2. This effect was not due to MNK's known functions as eukaryotic initiation factor (eIF) 4G binding partner or eIF4E(S209) kinase. Rather, MNK catalytic activity enabled viral IRES-mediated translation/host cell cytotoxicity through negative regulation of the Ser/Arg (SR)-rich protein kinase (SRPK). Our investigations suggest that SRPK activity is a major determinant of type 1 IRES competency, host cell cytotoxicity, and viral proliferation in infected cells.We are targeting unfettered enterovirus IRES activity in cancer with PVSRIPO, the type 1 live-attenuated poliovirus (PV) (Sabin) vaccine containing a human rhinovirus type 2 (HRV2) IRES. A phase I clinical trial of PVSRIPO with intratumoral inoculation in patients with recurrent glioblastoma (GBM) is showing early promise. Viral translation proficiency in infected GBM cells is a core requirement for the antineoplastic efficacy of PVSRIPO. Therefore, it is critically important to understand the mechanisms controlling viral cap-independent translation in infected host cells.

Authors
Brown, MC; Bryant, JD; Dobrikova, EY; Shveygert, M; Bradrick, SS; Chandramohan, V; Bigner, DD; Gromeier, M
MLA Citation
Brown, MC, Bryant, JD, Dobrikova, EY, Shveygert, M, Bradrick, SS, Chandramohan, V, Bigner, DD, and Gromeier, M. "Induction of viral, 7-methyl-guanosine cap-independent translation and oncolysis by mitogen-activated protein kinase-interacting kinase-mediated effects on the serine/arginine-rich protein kinase." Journal of virology 88.22 (November 2014): 13135-13148.
PMID
25187541
Source
epmc
Published In
Journal of virology
Volume
88
Issue
22
Publish Date
2014
Start Page
13135
End Page
13148
DOI
10.1128/jvi.01883-14

Mitogen-activated protein kinase-interacting kinase regulates mTOR/AKT signaling and controls the serine/arginine-rich protein kinase-responsive type 1 internal ribosome entry site-mediated translation and viral oncolysis.

Translation machinery is a major recipient of the principal mitogenic signaling networks involving Raf-ERK1/2 and phosphoinositol 3-kinase (PI3K)-mechanistic target of rapamycin (mTOR). Picornavirus internal ribosomal entry site (IRES)-mediated translation and cytopathogenic effects are susceptible to the status of such signaling cascades in host cells. We determined that tumor-specific cytotoxicity of the poliovirus/rhinovirus chimera PVSRIPO is facilitated by Raf-ERK1/2 signals to the mitogen-activated protein kinase (MAPK)-interacting kinase (MNK) and its effects on the partitioning/activity of the Ser/Arg (SR)-rich protein kinase (SRPK) (M. C. Brown, J. D. Bryant, E. Y. Dobrikova, M. Shveygert, S. S. Bradrick, V. Chandramohan, D. D. Bigner, and M, Gromeier, J. Virol. 22:13135-13148, 2014, doi:http://dx.doi.org/10.1128/JVI.01883-14). Here, we show that MNK regulates SRPK via mTOR and AKT. Our investigations revealed a MNK-controlled mechanism acting on mTORC2-AKT. The resulting suppression of AKT signaling attenuates SRPK activity to enhance picornavirus type 1 IRES translation and favor PVSRIPO tumor cell toxicity and killing.Oncolytic immunotherapy with PVSRIPO, the type 1 live-attenuated poliovirus (PV) (Sabin) vaccine containing a human rhinovirus type 2 (HRV2) IRES, is demonstrating early promise in clinical trials with intratumoral infusion in recurrent glioblastoma (GBM). Our investigations demonstrate that the core mechanistic principle of PVSRIPO, tumor-selective translation and cytotoxicity, relies on constitutive ERK1/2-MNK signals that counteract the deleterious effects of runaway AKT-SRPK activity in malignancy.

Authors
Brown, MC; Dobrikov, MI; Gromeier, M
MLA Citation
Brown, MC, Dobrikov, MI, and Gromeier, M. "Mitogen-activated protein kinase-interacting kinase regulates mTOR/AKT signaling and controls the serine/arginine-rich protein kinase-responsive type 1 internal ribosome entry site-mediated translation and viral oncolysis." Journal of virology 88.22 (November 2014): 13149-13160.
PMID
25187540
Source
epmc
Published In
Journal of virology
Volume
88
Issue
22
Publish Date
2014
Start Page
13149
End Page
13160
DOI
10.1128/jvi.01884-14

Mitotic phosphorylation of eukaryotic initiation factor 4G1 (eIF4G1) at Ser1232 by Cdk1:cyclin B inhibits eIF4A helicase complex binding with RNA.

During mitosis, global translation is suppressed, while synthesis of proteins with vital mitotic roles must go on. Prior evidence suggests that the mitotic translation shift involves control of initiation. Yet, no signals specifically targeting translation initiation factors during mitosis have been identified. We used phosphoproteomics to investigate the central translation initiation scaffold and "ribosome adaptor," eukaryotic initiation factor 4G1 (eIF4G1) in interphase or nocodazole-arrested mitotic cells. This approach and kinase inhibition assays, in vitro phosphorylation with recombinant kinase, and kinase depletion-reconstitution experiments revealed that Ser1232 in eIF4G1 is phosphorylated by cyclin-dependent kinase 1 (Cdk1):cyclin B during mitosis. Ser1232 is located in an unstructured region of the C-terminal portion of eIF4G1 that coordinates assembly of the eIF4G/-4A/-4B helicase complex and binding of the mitogen-activated protein kinase (MAPK) signal-integrating kinase, Mnk. Intense phosphorylation of Ser1232 in mitosis strongly enhanced the interactions of eIF4A with HEAT domain 2 of eIF4G and decreased association of eIF4G/-4A with RNA. Our findings implicate phosphorylation of eIF4G1(Ser1232) by Cdk1:cyclin B and its inhibitory effects on eIF4A helicase activity in the mitotic translation initiation shift.

Authors
Dobrikov, MI; Shveygert, M; Brown, MC; Gromeier, M
MLA Citation
Dobrikov, MI, Shveygert, M, Brown, MC, and Gromeier, M. "Mitotic phosphorylation of eukaryotic initiation factor 4G1 (eIF4G1) at Ser1232 by Cdk1:cyclin B inhibits eIF4A helicase complex binding with RNA." Mol Cell Biol 34.3 (February 2014): 439-451.
PMID
24248602
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
34
Issue
3
Publish Date
2014
Start Page
439
End Page
451
DOI
10.1128/MCB.01046-13

Oncolytic polio virotherapy of cancer

© 2014 American Cancer Society. Recently, the century-old idea of targeting cancer with viruses (oncolytic viruses) has come of age, and promise has been documented in early stage and several late-stage clinical trials in a variety of cancers. Although originally prized for their direct tumor cytotoxicity (oncolytic virotherapy), recently, the proinflammatory and immunogenic effects of viral tumor infection (oncolytic immunotherapy) have come into focus. Indeed, a capacity for eliciting broad, sustained antineoplastic effects stemming from combined direct viral cytotoxicity, innate antiviral activation, stromal proinflammatory stimulation, and recruitment of adaptive immune effector responses is the greatest asset of oncolytic viruses. However, it also is the source for enormous mechanistic complexity that must be considered for successful clinical translation. Because of fundamentally different relationships with their hosts (malignant or not), diverse replication strategies, and distinct modes of tumor cytotoxicity/killing, oncolytic viruses should not be referred to collectively. These agents must be evaluated based on their individual merits. In this review, the authors highlight key mechanistic principles of cancer treatment with the polio:rhinovirus chimera PVSRIPO and their implications for oncolytic immunotherapy in the clinic.

Authors
Brown, MC; Dobrikova, EY; Dobrikov, MI; Walton, RW; Gemberling, SL; Nair, SK; Desjardins, A; Sampson, JH; Friedman, HS; Friedman, AH; Tyler, DS; Bigner, DD; Gromeier, M
MLA Citation
Brown, MC, Dobrikova, EY, Dobrikov, MI, Walton, RW, Gemberling, SL, Nair, SK, Desjardins, A, Sampson, JH, Friedman, HS, Friedman, AH, Tyler, DS, Bigner, DD, and Gromeier, M. "Oncolytic polio virotherapy of cancer." Cancer 120.21 (January 1, 2014): 3277-3286. (Review)
Source
scopus
Published In
Cancer
Volume
120
Issue
21
Publish Date
2014
Start Page
3277
End Page
3286
DOI
10.1002/cncr.28862

Current immunotherapeutic targets in gliomas

The prospect for high-grade astrocytic tumor patients continues to be dismal, evenwith advances in surgery, radiotherapy, and chemotherapy. There is a need for thedevelopment of novel therapies that would eliminate the heterogeneous and diffuseastrocytic tumor cells.Innovative therapeutic approaches for the treatment of central nervous systemneoplasia, such as antibody-mediated immunotherapeutics, cellular immunotherapeutics,and oncolytic viruses targeting tumor-associated antigens, have emerged during the pastfew years.Herein we review several of the glioma-associated antigens and the current status ofimmunotherapeutics and oncolytic viruses targeting these antigens. © 2013 Nova Science Publishers, Inc. All rights reserved.

Authors
Chandramohan, V; Mitchell, DA; Gromeier, M; Sampson, JH; Bigner, DD
MLA Citation
Chandramohan, V, Mitchell, DA, Gromeier, M, Sampson, JH, and Bigner, DD. "Current immunotherapeutic targets in gliomas." (May 1, 2013): 287-305. (Chapter)
Source
scopus
Publish Date
2013
Start Page
287
End Page
305

Dynamic regulation of the translation initiation helicase complex by mitogenic signal transduction to eukaryotic translation initiation factor 4G.

Eukaryotic translation initiation factor 4F (eIF4F), comprising the cap-binding protein eIF4E, the helicase eIF4A, and the central scaffold eIF4G, is a convergence node for a complex signaling network that controls protein synthesis. Together with eIF3 and eIF4A/4B, eIF4G recruits ribosomal subunits to mRNAs and facilitates 5' untranslated region unwinding. Mammalian eIF4G contains three HEAT domains and unstructured regions involved in protein-protein interactions. Despite detailed eIF4G structure data, the mechanisms controlling initiation scaffold formation remain obscure. We found a new, highly regulated eIF4B/-3 binding site within the HEAT-1/-2 interdomain linker, harboring two phosphorylation sites that we identified as substrates for Erk1/2 and casein kinase 2. Phorbol ester-induced sequential phosphorylation of both sites detached HEAT-2 from the complex with eIF4A/-4B/-3 and stimulated the association of HEAT-3 with the mitogen-activated protein kinase signal integrating kinase Mnk1. Our results provide a mechanistic link between intracellular signal transduction and dynamic initiation complex formation coordinated by flexible eIF4G structure.

Authors
Dobrikov, MI; Dobrikova, EY; Gromeier, M
MLA Citation
Dobrikov, MI, Dobrikova, EY, and Gromeier, M. "Dynamic regulation of the translation initiation helicase complex by mitogenic signal transduction to eukaryotic translation initiation factor 4G." Mol Cell Biol 33.5 (March 2013): 937-946.
PMID
23263986
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
33
Issue
5
Publish Date
2013
Start Page
937
End Page
946
DOI
10.1128/MCB.01441-12

p38α mitogen-activated protein kinase depletion and repression of signal transduction to translation machinery by miR-124 and -128 in neurons.

The p38α to p38δ mitogen-activated protein kinases (MAPKs) are central regulatory nodes coordinating acute stress and inflammatory responses. Their activation leads to rapid adjustment of protein synthesis, for instance translational induction of proinflammatory cytokines. The only known direct link of p38 to translation machinery is the MAPK signal-integrating kinase Mnk. Only p38α and p38β transcripts are ubiquitously expressed. These mRNAs encode highly conserved proteins that equally phosphorylate recombinant Mnk1 in vitro. We discovered that expression of the p38α protein, but not the p38β isoform, is suppressed in the brain. This is due to p38α depletion by two neuron-selective microRNAs (miRNAs), miR-124 and -128. Suppression of p38α protein was reversed by miR-124/-128 antisense oligonucleotides in primary explant neuronal cultures. Targeted p38α depletion reduced Mnk1 activation, which cannot be compensated by p38β. Our research shows that p38α alone controls acute stress and cytokine signaling from p38 MAPK to translation machinery. This regulatory axis is greatly diminished in neurons, which may insulate brain physiology and function from p38α-Mnk1-mediated signaling.

Authors
Lawson, SK; Dobrikova, EY; Shveygert, M; Gromeier, M
MLA Citation
Lawson, SK, Dobrikova, EY, Shveygert, M, and Gromeier, M. "p38α mitogen-activated protein kinase depletion and repression of signal transduction to translation machinery by miR-124 and -128 in neurons." Mol Cell Biol 33.1 (January 2013): 127-135.
PMID
23109423
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
33
Issue
1
Publish Date
2013
Start Page
127
End Page
135
DOI
10.1128/MCB.00695-12

PD-0332991, a CDK4/6 inhibitor, significantly prolongs survival in a genetically engineered mouse model of brainstem glioma.

Diffuse intrinsic pontine glioma (DIPG) is an incurable tumor that arises in the brainstem of children. To date there is not a single approved drug to effectively treat these tumors and thus novel therapies are desperately needed. Recent studies suggest that a significant fraction of these tumors contain alterations in cell cycle regulatory genes including amplification of the D-type cyclins and CDK4/6, and less commonly, loss of Ink4a-ARF leading to aberrant cell proliferation. In this study, we evaluated the therapeutic approach of targeting the cyclin-CDK-Retinoblastoma (Rb) pathway in a genetically engineered PDGF-B-driven brainstem glioma (BSG) mouse model. We found that PD-0332991 (PD), a CDK4/6 inhibitor, induces cell-cycle arrest in our PDGF-B; Ink4a-ARF deficient model both in vitro and in vivo. By contrast, the PDGF-B; p53 deficient model was mostly resistant to treatment with PD. We noted that a 7-day treatment course with PD significantly prolonged survival by 12% in the PDGF-B; Ink4a-ARF deficient BSG model. Furthermore, a single dose of 10 Gy radiation therapy (RT) followed by 7 days of treatment with PD increased the survival by 19% in comparison to RT alone. These findings provide the rationale for evaluating PD in children with Ink4a-ARF deficient gliomas.

Authors
Barton, KL; Misuraca, K; Cordero, F; Dobrikova, E; Min, HD; Gromeier, M; Kirsch, DG; Becher, OJ
MLA Citation
Barton, KL, Misuraca, K, Cordero, F, Dobrikova, E, Min, HD, Gromeier, M, Kirsch, DG, and Becher, OJ. "PD-0332991, a CDK4/6 inhibitor, significantly prolongs survival in a genetically engineered mouse model of brainstem glioma. (Published online)" PLoS One 8.10 (2013): e77639-.
PMID
24098593
Source
pubmed
Published In
PloS one
Volume
8
Issue
10
Publish Date
2013
Start Page
e77639
DOI
10.1371/journal.pone.0077639

Attenuation of neurovirulence, biodistribution, and shedding of a poliovirus:rhinovirus chimera after intrathalamic inoculation in Macaca fascicularis.

A dependence of poliovirus on an unorthodox translation initiation mode can be targeted selectively to drive viral protein synthesis and cytotoxicity in malignant cells. Transformed cells are naturally susceptible to poliovirus, due to widespread ectopic upregulation of the poliovirus receptor, Necl-5, in ectodermal/neuroectodermal cancers. Viral tumor cell killing and the host immunologic response it engenders produce potent, lasting antineoplastic effects in animal tumor models. Clinical application of this principle depends on unequivocal demonstration of safety in primate models for paralytic poliomyelitis. We conducted extensive dose-range-finding, toxicity, biodistribution, shedding, and neutralizing antibody studies of the prototype oncolytic poliovirus recombinant, PVS-RIPO, after intrathalamic inoculation in Macaca fascicularis. These studies suggest that intracerebral PVS-RIPO inoculation does not lead to viral propagation in the central nervous system (CNS), does not cause histopathological CNS lesions or neurological symptoms that can be attributed to the virus, is not associated with extraneural virus dissemination or replication and does not induce shedding of virus with stool. Intrathalamic PVS-RIPO inoculation induced neutralizing antibody responses against poliovirus serotype 1 in all animals studied.

Authors
Dobrikova, EY; Goetz, C; Walters, RW; Lawson, SK; Peggins, JO; Muszynski, K; Ruppel, S; Poole, K; Giardina, SL; Vela, EM; Estep, JE; Gromeier, M
MLA Citation
Dobrikova, EY, Goetz, C, Walters, RW, Lawson, SK, Peggins, JO, Muszynski, K, Ruppel, S, Poole, K, Giardina, SL, Vela, EM, Estep, JE, and Gromeier, M. "Attenuation of neurovirulence, biodistribution, and shedding of a poliovirus:rhinovirus chimera after intrathalamic inoculation in Macaca fascicularis." J Virol 86.5 (March 2012): 2750-2759.
PMID
22171271
Source
pubmed
Published In
Journal of virology
Volume
86
Issue
5
Publish Date
2012
Start Page
2750
End Page
2759
DOI
10.1128/JVI.06427-11

Oncolytic poliovirus against malignant glioma.

In cancerous cells, physiologically tight regulation of protein synthesis is lost, contributing to uncontrolled growth and proliferation. We describe a novel experimental cancer therapy approach based on genetically recombinant poliovirus that targets an intriguing aberration of translation control in malignancy. This strategy is based on the confluence of several factors enabling specific and efficacious cancer cell targeting. Poliovirus naturally targets the vast majority of ectodermal/neuroectodermal cancers expressing its cellular receptor. Evidence from glioblastoma patients suggests that the poliovirus receptor is ectopically upregulated on tumor cells and may be associated with stem cell-like cancer cell populations and proliferating tumor vasculature. We exploit poliovirus' reliance on an unorthodox mechanism of protein synthesis initiation to selectively drive viral translation, propagation and cytotoxicity in glioblastoma. PVSRIPO, a prototype nonpathogenic poliovirus recombinant, is scheduled to enter clinical investigation against glioblastoma.

Authors
Goetz, C; Dobrikova, E; Shveygert, M; Dobrikov, M; Gromeier, M
MLA Citation
Goetz, C, Dobrikova, E, Shveygert, M, Dobrikov, M, and Gromeier, M. "Oncolytic poliovirus against malignant glioma." Future Virol 6.9 (September 2011): 1045-1058.
PMID
21984883
Source
pubmed
Published In
Future virology
Volume
6
Issue
9
Publish Date
2011
Start Page
1045
End Page
1058
DOI
10.2217/fvl.11.76

Phosphorylation of eukaryotic translation initiation factor 4G1 (eIF4G1) by protein kinase C{alpha} regulates eIF4G1 binding to Mnk1.

Signal transduction through mitogen-activated protein kinases (MAPKs) is implicated in growth and proliferation control through translation regulation and involves posttranslational modification of translation initiation factors. For example, convergent MAPK signals to Mnk1 lead to phosphorylation of eukaryotic translation initiation factor 4E (eIF4E), which has been linked to malignant transformation. However, understanding the compound effects of mitogenic signaling on the translation apparatus and on protein synthesis control remains elusive. This is particularly true for the central scaffold of the translation initiation apparatus and ribosome adaptor eIF4G. To unravel the effects of signal transduction to eIF4G on translation, we used specific activation of protein kinase C (PKC)-Ras-Erk signaling with phorbol esters. Phospho-proteomic and mutational analyses revealed that eIF4G1 is a substrate for PKCα at Ser1186. We show that PKCα activation elicits a cascade of orchestrated phosphorylation events that may modulate eIF4G1 structure and control interaction with the eIF4E kinase, Mnk1.

Authors
Dobrikov, M; Dobrikova, E; Shveygert, M; Gromeier, M
MLA Citation
Dobrikov, M, Dobrikova, E, Shveygert, M, and Gromeier, M. "Phosphorylation of eukaryotic translation initiation factor 4G1 (eIF4G1) by protein kinase C{alpha} regulates eIF4G1 binding to Mnk1." Mol Cell Biol 31.14 (July 2011): 2947-2959.
PMID
21576361
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
31
Issue
14
Publish Date
2011
Start Page
2947
End Page
2959
DOI
10.1128/MCB.05589-11

Adaptation of an ICAM-1-tropic enterovirus to the mouse respiratory tract.

Respiratory tract (RT) infections by members of the enterovirus (EV) genus of the Picornaviridae family are the most frequent cause for the common cold and a major factor in the exacerbation of chronic pulmonary diseases. The lack of a practical small-animal model for these infections has obstructed insight into pathogenic mechanisms of the common cold and their role in chronic RT illness and has hampered preclinical evaluation of antiviral strategies. Despite significant efforts, it has been difficult to devise rodent models that exhibit viral replication in the RT. This is due mainly to well-known intracellular host restrictions of EVs with RT tropism in rodent cells. We report the evolution of variants of the common-cold-causing coxsackievirus A21, an EV with tropism for the human intercellular adhesion molecule 1 (hICAM-1), through serial passage in the lungs of mice transgenic for the hICAM-1 gene. This process was accompanied by multiple changes in the viral genome, suggesting exquisite adaptation of hICAM-1-tropic enteroviruses to the specific growth conditions within the RT. In vivo mouse RT-adapted, variant coxsackievirus A21 exhibited replication competence in the lungs of hICAM-1 transgenic mice, providing a basis for unraveling EV-host interactions in the mouse RT.

Authors
Wang, ES; Dobrikova, E; Goetz, C; Dufresne, AT; Gromeier, M
MLA Citation
Wang, ES, Dobrikova, E, Goetz, C, Dufresne, AT, and Gromeier, M. "Adaptation of an ICAM-1-tropic enterovirus to the mouse respiratory tract." J Virol 85.11 (June 2011): 5606-5617.
PMID
21450825
Source
pubmed
Published In
Journal of virology
Volume
85
Issue
11
Publish Date
2011
Start Page
5606
End Page
5617
DOI
10.1128/JVI.01502-10

MAPK signal-integrating kinase controls cap-independent translation and cell type-specific cytotoxicity of an oncolytic poliovirus.

Many animal viruses exhibit proficient growth in transformed cells, a property that has been harnessed for the development of novel therapies against cancer. Despite overwhelming evidence for this phenomenon, understanding of the molecular mechanisms enabling tumor-cell killing is rudimentary for most viruses. We report here that growth and cytotoxicity of the prototype oncolytic poliovirus (PV), PVSRIPO, in glioblastoma multiforme (GBM) is promoted by mitogen-activated protein kinases (MAPKs) converging on the MAPK signal-integrating kinase 1 (Mnk1) and its primary substrate, the eukaryotic initiation factor (eIF) 4E. Inducing Mnk1-catalyzed eIF4E phosphorylation through expression of oncogenic Ras substantially enhanced PVSRIPO translation, replication, and cytotoxicity in resistant cells. This effect was mimicked by expression of constitutively active forms of Mnk1 and correlated with enhanced translation of subgenomic reporter RNAs. Our findings implicate Mnk1 activity in stimulation of PVSRIPO cap-independent translation, an effect that can be synergistically enhanced by inhibition of the phosphoinositide-3 kinase (PI3K).

Authors
Goetz, C; Everson, RG; Zhang, LC; Gromeier, M
MLA Citation
Goetz, C, Everson, RG, Zhang, LC, and Gromeier, M. "MAPK signal-integrating kinase controls cap-independent translation and cell type-specific cytotoxicity of an oncolytic poliovirus." Mol Ther 18.11 (November 2010): 1937-1946.
PMID
20648000
Source
pubmed
Published In
Molecular Therapy
Volume
18
Issue
11
Publish Date
2010
Start Page
1937
End Page
1946
DOI
10.1038/mt.2010.145

Regulation of eukaryotic initiation factor 4E (eIF4E) phosphorylation by mitogen-activated protein kinase occurs through modulation of Mnk1-eIF4G interaction.

The m(7)G cap binding protein eukaryotic initiation factor 4E (eIF4E) is a rate-limiting determinant of protein synthesis. Elevated eIF4E levels, commonly associated with neoplasia, promote oncogenesis, and phosphorylation of eIF4E at Ser209 is critical for its tumorigenic potential. eIF4E phosphorylation is catalyzed by mitogen-activated protein kinase (MAPK)-interacting serine/threonine kinase (Mnk), a substrate of Erk1/2 and p38 MAPKs. Interaction with the scaffolding protein eIF4G, which also binds eIF4E, brings Mnk and its substrate into physical proximity. Thus, Mnk-eIF4G interaction is important for eIF4E phosphorylation. Through coimmunoprecipitation assays, we showed that MAPK-mediated phosphorylation of the Mnk1 active site controls eIF4G binding. Utilizing a naturally occurring splice variant, we demonstrated that the C-terminal domain of Mnk1 restricts its interaction with eIF4G, preventing eIF4E phosphorylation in the absence of MAPK signaling. Furthermore, using a small-molecule Mnk1 inhibitor and kinase-dead mutant, we established that Mnk1 autoregulates its interaction with eIF4G, releasing itself from the scaffold after phosphorylation of its substrate. Our findings indicate tight control of eIF4E phosphorylation through modulation of Mnk1-eIF4G interaction.

Authors
Shveygert, M; Kaiser, C; Bradrick, SS; Gromeier, M
MLA Citation
Shveygert, M, Kaiser, C, Bradrick, SS, and Gromeier, M. "Regulation of eukaryotic initiation factor 4E (eIF4E) phosphorylation by mitogen-activated protein kinase occurs through modulation of Mnk1-eIF4G interaction." Mol Cell Biol 30.21 (November 2010): 5160-5167.
PMID
20823271
Source
pubmed
Published In
Molecular and Cellular Biology
Volume
30
Issue
21
Publish Date
2010
Start Page
5160
End Page
5167
DOI
10.1128/MCB.00448-10

Tissue type-specific expression of the dsRNA-binding protein 76 and genome-wide elucidation of its target mRNAs.

BACKGROUND: RNA-binding proteins accompany all steps in the life of mRNAs and provide dynamic gene regulatory functions for rapid adjustment to changing extra- or intracellular conditions. The association of RNA-binding proteins with their targets is regulated through changing subcellular distribution, post-translational modification or association with other proteins. METHODOLOGY: We demonstrate that the dsRNA binding protein 76 (DRBP76), synonymous with nuclear factor 90, displays inherently distinct tissue type-specific subcellular distribution in the normal human central nervous system and in malignant brain tumors of glial origin. Altered subcellular localization and isoform distribution in malignant glioma indicate that tumor-specific changes in DRBP76-related gene products and their regulatory functions may contribute to the formation and/or maintenance of these tumors. To identify endogenous mRNA targets of DRBP76, we performed RNA-immunoprecipitation and genome-wide microarray analyses in HEK293 cells, and identified specific classes of transcripts encoding critical functions in cellular metabolism. SIGNIFICANCE: Our data suggest that physiologic DRBP76 expression, isoform distribution and subcellular localization are profoundly altered upon malignant transformation. Thus, the functional role of DRBP76 in co- or post-transcriptional gene regulation may contribute to the neoplastic phenotype.

Authors
Neplioueva, V; Dobrikova, EY; Mukherjee, N; Keene, JD; Gromeier, M
MLA Citation
Neplioueva, V, Dobrikova, EY, Mukherjee, N, Keene, JD, and Gromeier, M. "Tissue type-specific expression of the dsRNA-binding protein 76 and genome-wide elucidation of its target mRNAs. (Published online)" PLoS One 5.7 (July 23, 2010): e11710-.
Website
http://hdl.handle.net/10161/4556
PMID
20668518
Source
pubmed
Published In
PloS one
Volume
5
Issue
7
Publish Date
2010
Start Page
e11710
DOI
10.1371/journal.pone.0011710

Preparing an oncolytic poliovirus recombinant for clinical application against glioblastoma multiforme.

PVS-RIPO is a genetically recombinant, non-pathogenic poliovirus chimera with a tumor-specific conditional replication phenotype. Consisting of the genome of the live attenuated poliovirus type 1 (Sabin) vaccine with its cognate IRES element replaced with that of human rhinovirus type 2, PVS-RIPO displays an inability to translate its genome in untransformed neuronal cells, but effectively does so in cells originating from primary tumors in the central nervous system or other cancers. Hence, PVS-RIPO unleashes potent cytotoxic effects on infected cancer cells and produces sustained anti-tumoral responses in animal tumor models. PVS-RIPO presents a novel approach to the treatment of patients with glioblastoma multiforme, based on conditions favoring an unconventional viral translation initiation mechanism in cancerous cells. In this review we summarize advances in the understanding of major molecular determinants of PVS-RIPO oncolytic efficacy and safety and discuss their implications for upcoming clinical investigations.

Authors
Goetz, C; Gromeier, M
MLA Citation
Goetz, C, and Gromeier, M. "Preparing an oncolytic poliovirus recombinant for clinical application against glioblastoma multiforme." Cytokine Growth Factor Rev 21.2-3 (April 2010): 197-203. (Review)
PMID
20299272
Source
pubmed
Published In
Cytokine and Growth Factor Reviews
Volume
21
Issue
2-3
Publish Date
2010
Start Page
197
End Page
203
DOI
10.1016/j.cytogfr.2010.02.005

Poly(A)-binding protein modulates mRNA susceptibility to cap-dependent miRNA-mediated repression.

MicroRNAs (miRNAs) regulate gene expression post-transcriptionally through binding specific sites within the 3' untranslated regions (UTRs) of their target mRNAs. Numerous investigations have documented repressive effects of miRNAs and identified factors required for their activity. However, the precise mechanisms by which miRNAs modulate gene expression are still obscure. Here, we have examined the effects of multiple miRNAs on diverse target transcripts containing artificial or naturally occurring 3' UTRs in human cell culture. In agreement with previous studies, we report that both the 5' m(7)G cap and 3' poly(A) tail are essential for maximum miRNA repression. These cis-acting elements also conferred miRNA susceptibility to target mRNAs translating under the control of viral- and eukaryotic mRNA-derived 5' UTR structures that enable cap-independent translation. Additionally, we evaluated a role for the poly(A)-binding protein (PABP) in miRNA function utilizing multiple approaches to modulate levels of active PABP in cells. PABP expression and activity inversely correlated with the strength of miRNA silencing, in part due to antagonism of target mRNA deadenylation. Together, these findings further define the cis- and trans-acting factors that modulate miRNA efficacy.

Authors
Walters, RW; Bradrick, SS; Gromeier, M
MLA Citation
Walters, RW, Bradrick, SS, and Gromeier, M. "Poly(A)-binding protein modulates mRNA susceptibility to cap-dependent miRNA-mediated repression." RNA 16.1 (January 2010): 239-250.
PMID
19934229
Source
pubmed
Published In
RNA (New York, N.Y.)
Volume
16
Issue
1
Publish Date
2010
Start Page
239
End Page
250
DOI
10.1261/rna.1795410

Herpes simplex virus proteins ICP27 and UL47 associate with polyadenylate-binding protein and control its subcellular distribution.

Human pathogenic viruses manipulate host cell translation machinery to ensure efficient expression of viral genes and to thwart host cell protein synthesis. Viral strategies include cleaving translation factors, manipulating translation factor abundance and recruitment into translation initiation complexes, or expressing viral translation factor analogs. Analyzing translation factors in herpes simplex virus type 1 (HSV-1)-infected HeLa cells, we found diminished association of the polyadenylate-binding protein (PABP) with the cap-binding complex. Although total PABP levels were unchanged, HSV-1 infection prompted accumulation of cytoplasmic PABPC1, but not its physiologic binding partner PABP-interacting protein 2 (Paip2), in the nucleus. Using glutathione S-transferase-PABP pull-down and proteomic analyses, we identified several viral proteins interacting with PABPC1 including tegument protein UL47 and infected-cell protein ICP27. Transient expression of ICP27 and UL47 in HeLa cells suggested that ICP27 and UL47 jointly displace Paip2 from PABP. ICP27 expression alone was sufficient to cause PABPC1 redistribution to the nucleus. ICP27 and UL47 did not alter translation efficiency of transfected reporter RNAs but modulated transcript abundance and expression of reporter cDNAs in transfected cells. This indicates that redistribution of PABPC1 may be involved in co- and posttranscriptional regulation of mRNA processing and/or nuclear export by HSV-1 gene regulatory proteins.

Authors
Dobrikova, E; Shveygert, M; Walters, R; Gromeier, M
MLA Citation
Dobrikova, E, Shveygert, M, Walters, R, and Gromeier, M. "Herpes simplex virus proteins ICP27 and UL47 associate with polyadenylate-binding protein and control its subcellular distribution." J Virol 84.1 (January 2010): 270-279.
PMID
19864386
Source
pubmed
Published In
Journal of virology
Volume
84
Issue
1
Publish Date
2010
Start Page
270
End Page
279
DOI
10.1128/JVI.01740-09

Identification of gemin5 as a novel 7-methylguanosine cap-binding protein.

BACKGROUND: A unique attribute of RNA molecules synthesized by RNA polymerase II is the presence of a 7-methylguanosine (m(7)G) cap structure added co-transcriptionally to the 5' end. Through its association with trans-acting effector proteins, the m(7)G cap participates in multiple aspects of RNA metabolism including localization, translation and decay. However, at present relatively few eukaryotic proteins have been identified as factors capable of direct association with m(7)G. METHODOLOGY/PRINCIPAL FINDINGS: Employing an unbiased proteomic approach, we identified gemin5, a component of the survival of motor neuron (SMN) complex, as a factor capable of direct and specific interaction with the m(7)G cap. Gemin5 was readily purified by cap-affinity chromatography in contrast to other SMN complex proteins. Investigating the underlying basis for this observation, we found that purified gemin5 associates with m(7)G-linked sepharose in the absence of detectable eIF4E, and specifically crosslinks to radiolabeled cap structure after UV irradiation. Deletion analysis revealed that an intact set of WD repeat domains located in the N-terminal half of gemin5 are required for cap-binding. Moreover, using structural modeling and site-directed mutagenesis, we identified two proximal aromatic residues located within the WD repeat region that significantly impact m(7)G association. CONCLUSIONS/SIGNIFICANCE: This study rigorously identifies gemin5 as a novel cap-binding protein and describes an unprecedented role for WD repeat domains in m(7)G recognition. The findings presented here will facilitate understanding of gemin5's role in the metabolism of non-coding snRNAs and perhaps other RNA pol II transcripts.

Authors
Bradrick, SS; Gromeier, M
MLA Citation
Bradrick, SS, and Gromeier, M. "Identification of gemin5 as a novel 7-methylguanosine cap-binding protein. (Published online)" PLoS One 4.9 (September 14, 2009): e7030-.
PMID
19750007
Source
pubmed
Published In
PloS one
Volume
4
Issue
9
Publish Date
2009
Start Page
e7030
DOI
10.1371/journal.pone.0007030

Evaluation of IRES-mediated, cell-type-specific cytotoxicity of poliovirus using a colorimetric cell proliferation assay.

PVS-RIPO is a recombinant oncolytic poliovirus designed for clinical application to target CD155 expressing malignant gliomas and other malignant diseases. PVS-RIPO does not replicate in healthy neurons and is therefore non-pathogenic in rodent and non-human primate models of poliomyelitis. A tetrazolium salt dye-based cellular assay was developed and qualified to define the cytotoxicity of virus preparations on susceptible cells and to explore the target cell specificity of PVS-RIPO. In this assay, PVS-RIPO inhibited proliferation of U87-MG astrocytoma cells in a dose-dependent manner. However, HEK293 cells were much less susceptible to cell killing by PVS-RIPO. In contrast, the Sabin type 1 live attenuated poliovirus vaccine strain (PV(1)S) was cytotoxic to both HEK293 and U87-MG cells. The correlation between expression of CD155 and cytotoxicity was also explored using six different cell lines. There was little or no expression of CD155 and PVS-RIPO-induced cytotoxicity in Jurkat and Daudi cells. HEK293 was the only cell line tested that showed CD155 expression and resistance to PVS-RIPO cytotoxicity. The results indicate that differential cytotoxicity measured by the colorimetric assay can be used to evaluate the cytotoxicity and cell-type specificity of recombinant strains of poliovirus and to demonstrate lot to lot consistency during the manufacture of viruses intended for clinical use.

Authors
Yang, X; Chen, E; Jiang, H; Muszynski, K; Harris, RD; Giardina, SL; Gromeier, M; Mitra, G; Soman, G
MLA Citation
Yang, X, Chen, E, Jiang, H, Muszynski, K, Harris, RD, Giardina, SL, Gromeier, M, Mitra, G, and Soman, G. "Evaluation of IRES-mediated, cell-type-specific cytotoxicity of poliovirus using a colorimetric cell proliferation assay." J Virol Methods 155.1 (January 2009): 44-54.
PMID
18951922
Source
pubmed
Published In
Journal of Virological Methods
Volume
155
Issue
1
Publish Date
2009
Start Page
44
End Page
54
DOI
10.1016/j.jviromet.2008.09.020

Recombinant oncolytic poliovirus eliminates glioma in vivo without genetic adaptation to a pathogenic phenotype.

Many viruses, either naturally occurring or as a result of genetic manipulation, exhibit conditional replication in transformed cells. This principle is the basis for experimental therapeutic approaches exploiting the oncolytic potential of such agents without the danger of collateral damage to resistant normal tissues. One of the potential obstacles to these approaches is the possibility of genetic adaptation of oncolytic viruses upon replication in susceptible tumor tissues. Genetic variation can reverse genetic manipulations of parental viral genomes that determine attenuation of virulence, selective tumor cell tropism or other desirable traits. Alternatively, it may convey new properties not originally associated with parental strains, e.g., adaptation to a human host range. We examined genetic stability of an oncolytic nonpathogenic poliovirus recombinant considered for therapy of recurrent glioblastoma multiforme (GBM). This was done by serial passage experiments in glioma xenografts in vivo and investigation of phenotypic and genotypic markers of attenuation. Intratumoral inoculation of oncolytic poliovirus produced efficient tumor regress and elimination without altering temperature-sensitive growth, selective cytotoxicity, or genetic markers of attenuation of virus recovered from inoculated animals. Our studies demonstrate that active viral oncolysis of malignant glioma does not alter the conditional replication properties of oncolytic nonpathogenic poliovirus recombinants.

Authors
Dobrikova, EY; Broadt, T; Poiley-Nelson, J; Yang, X; Soman, G; Giardina, S; Harris, R; Gromeier, M
MLA Citation
Dobrikova, EY, Broadt, T, Poiley-Nelson, J, Yang, X, Soman, G, Giardina, S, Harris, R, and Gromeier, M. "Recombinant oncolytic poliovirus eliminates glioma in vivo without genetic adaptation to a pathogenic phenotype." Mol Ther 16.11 (November 2008): 1865-1872.
PMID
18766173
Source
pubmed
Published In
Molecular Therapy
Volume
16
Issue
11
Publish Date
2008
Start Page
1865
End Page
1872
DOI
10.1038/mt.2008.184

Activation of cap-independent translation by variant eukaryotic initiation factor 4G in vivo.

Protein synthesis is tightly controlled by assembly of an intricate ribonucleoprotein complex at the m(7)GTP-cap on eukaryotic mRNAs. Ensuing linear scanning of the 5' untranslated region (UTR) is believed to transfer the preinitiation complex to the initiation codon. Eukaryotic mRNAs are characterized by significant 5' UTR heterogeneity, raising the possibility of differential control of translation initiation rate at individual mRNAs. Curiously, many mRNAs with unconventional, highly structured 5' UTRs encode proteins with central biological roles in growth control, metabolism, or stress response. The 5' UTRs of such mRNAs may influence protein synthesis rate in multiple ways, but most significantly they have been implicated in mediating alternative means of translation initiation. Cap-independent initiation bypasses strict control over the formation of initiation intermediates at the m(7)GTP cap. However, the molecular mechanisms that favor alternative means of ribosome recruitment are not understood. Here we provide evidence that eukaryotic initiation factor (eIF) 4G controls cap-independent translation initiation at the c-myc and vascular endothelial growth factor (VEGF) 5' UTRs in vivo. Cap-independent translation was investigated in tetracycline-inducible cell lines expressing either full-length eIF4G or a C-terminal fragment (Ct) lacking interaction with eIF4E and poly(A) binding protein. Expression of Ct, but not intact eIF4G, potently stimulated cap-independent initiation at the c-myc/VEGF 5' UTRs. In vitro RNA-binding assays suggest that stimulation of cap-independent translation initiation by Ct is due to direct association with the c-myc/VEGF 5' UTR, enabling 43S preinitiation complex recruitment. Our work demonstrates that variant translation initiation factors enable unconventional translation initiation at mRNA subsets with distinct structural features.

Authors
Kaiser, C; Dobrikova, EY; Bradrick, SS; Shveygert, M; Herbert, JT; Gromeier, M
MLA Citation
Kaiser, C, Dobrikova, EY, Bradrick, SS, Shveygert, M, Herbert, JT, and Gromeier, M. "Activation of cap-independent translation by variant eukaryotic initiation factor 4G in vivo." RNA 14.10 (October 2008): 2170-2182.
PMID
18755839
Source
pubmed
Published In
RNA (New York, N.Y.)
Volume
14
Issue
10
Publish Date
2008
Start Page
2170
End Page
2182
DOI
10.1261/rna.1171808

Immunotherapy against angiogenesis-associated targets: evidence and implications for the treatment of malignant glioma.

Angiogenesis, the growth of new blood vessels from previously existing vasculature, is a requirement for tumor growth and metastasis. The first US FDA-approved drugs targeting angiogenesis have shown potential in the treatment of malignant gliomas. Immunotherapy as a treatment modality lends itself well to specifically targeting angiogenesis in tumors and may represent a powerful tool in the treatment of malignant gliomas. This review focuses on developments in immunotherapy targeting angiogenesis and tumor-vascular-specific endothelial cells using a variety of immunotherapeutic strategies including monoclonal antibodies and conjugated immunotoxins, as well as cellular, peptide, DNA and dendritic cell vaccines.

Authors
Everson, RG; Graner, MW; Gromeier, M; Vredenburgh, JJ; Desjardins, A; Reardon, DA; Friedman, HS; Friedman, AH; Bigner, DD; Sampson, JH
MLA Citation
Everson, RG, Graner, MW, Gromeier, M, Vredenburgh, JJ, Desjardins, A, Reardon, DA, Friedman, HS, Friedman, AH, Bigner, DD, and Sampson, JH. "Immunotherapy against angiogenesis-associated targets: evidence and implications for the treatment of malignant glioma." Expert Rev Anticancer Ther 8.5 (May 2008): 717-732. (Review)
PMID
18471045
Source
pubmed
Published In
Expert Review of Anticancer Therapy
Volume
8
Issue
5
Publish Date
2008
Start Page
717
End Page
732
DOI
10.1586/14737140.8.5.717

Molecular strategies for the treatment of malignant glioma--genes, viruses, and vaccines.

The standard treatment paradigm of surgery, radiation, and chemotherapy for malignant gliomas has only a modest effect on survival. It is well emphasized in the literature that despite aggressive multimodal therapy, most patients survive approximately 1 year after diagnosis, and less than 10% survive beyond 2 years. This dismal prognosis provides the impetus for ongoing investigations in search of improved therapeutics. Standard multimodal therapy has largely reached a plateau in terms of effectiveness, and there is now a growing body of literature on novel molecular approaches for the treatment of malignant gliomas. Gene therapy, oncolytic virotherapy, and immunotherapy are the major investigational approaches that have demonstrated promise in preclinical and early clinical studies. These new molecular technologies each have distinct advantages and limitations, and none has yet demonstrated a significant survival benefit in a phase II or III clinical trial. Molecular approaches may not lead to the discovery of a "magic bullet" for these aggressive tumors, but they may ultimately prove synergistic with more conventional approaches and lead to a broadening of the multimodal approach that is the current standard of care. This review will discuss the scientific background, therapeutic potential, and clinical limitations of these novel strategies with a focus on those that have made it to clinical trials.

Authors
Selznick, LA; Shamji, MF; Fecci, P; Gromeier, M; Friedman, AH; Sampson, J
MLA Citation
Selznick, LA, Shamji, MF, Fecci, P, Gromeier, M, Friedman, AH, and Sampson, J. "Molecular strategies for the treatment of malignant glioma--genes, viruses, and vaccines." Neurosurg Rev 31.2 (April 2008): 141-155. (Review)
PMID
18259789
Source
pubmed
Published In
Neurosurgical Review
Volume
31
Issue
2
Publish Date
2008
Start Page
141
End Page
155
DOI
10.1007/s10143-008-0121-0

Growth phenotypes and biosafety profiles in poliovirus-receptor transgenic mice of recombinant oncolytic polio/human rhinoviruses.

The use of oncolytic recombinant polioviruses has an important therapeutic potential in the treatment of human gliomas. This study was carried out to assess parameters of the utility of the oncolytic poliovirus/human rhinovirus type 2 chimeras (PV/HRV2). The prototype PV/HRV2 chimera was constructed containing the complete genome of wild-type PV type 1 (Mahoney) [PV1(M)] in which the cognate IRES was replaced with that of HRV2 [called PV1(RIPO)]. A derivative of PV1(RIPO) is PV1(RIPOS) in which the capsid coding region (P1) was replaced with the capsid-coding region of the PV type 1 (Sabin) [PV1(S)] vaccine strain. In addition, a third PV/HRV2 chimera was constructed containing the complete genome of PV1(S) in which the cognate IRES was replaced with that of HRV2 [termed PVS(RIPO)]. To analyze the growth phenotypes of PV/HRV2 recombinants [PV1(RIPO), PV1(RIPOS), PVS(RIPO)], one-step growth experiments were performed in four human cell lines at three different temperatures. To address the safety profile, PVS(RIPO) was injected into the brain of CD155 tg mice at the dose 10(7) PFU. Then, clinical signs, persistence of the virus in the CNS and genetic stability of PVS(RIPO) replicating in the CNS were evaluated. The data obtained in the present study suggest (i) a correlation between temperature-sensitive (ts) phenotype in both neuronal and non-neuronal cell lines and neuroattenuation in experimental animals, (ii) that PVS (RIPO) is genetically stable on replication in the CNS of poliovirus-susceptible mice. These findings highlight the safety of intracerebral inoculation of PVS(RIPO) for the treatment of human glioma.

Authors
Cello, J; Toyoda, H; Dejesus, N; Dobrikova, EY; Gromeier, M; Wimmer, E
MLA Citation
Cello, J, Toyoda, H, Dejesus, N, Dobrikova, EY, Gromeier, M, and Wimmer, E. "Growth phenotypes and biosafety profiles in poliovirus-receptor transgenic mice of recombinant oncolytic polio/human rhinoviruses." J Med Virol 80.2 (February 2008): 352-359.
PMID
18098139
Source
pubmed
Published In
Journal of Medical Virology
Volume
80
Issue
2
Publish Date
2008
Start Page
352
End Page
359
DOI
10.1002/jmv.21063

Poly(A)-binding protein is differentially required for translation mediated by viral internal ribosome entry sites.

The 3' poly(A) tail present on the majority of mature eukaryotic mRNAs is an important regulator of protein synthesis and mRNA stability. The poly(A) tail improves the efficiency of translation initiation through recruitment of PABP, enabling its interaction with eIF4F located at the mRNA 5'-end. Recent evidence has also implicated a possible role for PABP and the poly(A) tract in translation control at steps beyond the initiation phase. Similar to conventional mRNAs, plus-strand RNA virus genomes that utilize internal ribosome entry sites (IRESes) to promote cap-independent translation are influenced by PABP and poly(A) status. However, the relative contribution of these factors to translation initiation mediated by distinct IRESes is unclear. We have investigated cis- and trans-acting effects of poly(A) and PABP, respectively, on RNAs harboring IRESes from three diverse viruses: encephalomyocarditis virus (EMCV), hepatitis C virus (HCV), and coxsackievirus B3 (CBV3). A 3' poly(A) tract enhanced translation of both capped and IRES-containing reporter RNAs. However, only CBV3 and capped transcripts were stabilized as a result of polyadenylation. Correspondingly, translation of polyadenylated CBV3 and capped RNAs displayed heightened sensitivity to the PABP inhibitor Paip2 compared with EMCV and HCV. Sucrose density gradient analyses suggested a stimulatory role for PABP and 3' poly(A) in the CBV3 initiation phase, while assembly of HCV and EMCV RNAs into ribosomal complexes was little affected by either factor. Collectively, the observed differential effects of PABP and poly(A) on translation imply mechanistic differences between viral IRES elements and suggest modulating roles for PABP and the poly(A) tail at multiple phases of translation.

Authors
Bradrick, SS; Dobrikova, EY; Kaiser, C; Shveygert, M; Gromeier, M
MLA Citation
Bradrick, SS, Dobrikova, EY, Kaiser, C, Shveygert, M, and Gromeier, M. "Poly(A)-binding protein is differentially required for translation mediated by viral internal ribosome entry sites." RNA 13.9 (September 2007): 1582-1593.
PMID
17652408
Source
pubmed
Published In
RNA (New York, N.Y.)
Volume
13
Issue
9
Publish Date
2007
Start Page
1582
End Page
1593
DOI
10.1261/rna.556107

Genetic adaptation to untranslated region-mediated enterovirus growth deficits by mutations in the nonstructural proteins 3AB and 3CD.

Both untranslated regions (UTRs) of plus-strand RNA virus genomes jointly control translation and replication of viral genomes. In the case of the Enterovirus genus of the Picornaviridae family, the 5'UTR consists of a cloverleaf-like terminus preceding the internal ribosomal entry site (IRES) and the 3' terminus is composed of a structured 3'UTR and poly(A). The IRES and poly(A) have been implicated in translation control, and all UTR structures, in addition to cis-acting genetic elements mapping to the open reading frame, have been assigned roles in RNA replication. Viral UTRs are recognized by viral and host cell RNA-binding proteins that may co-determine genome stability, translation, plus- and minus-strand RNA replication, and scaffolding of viral replication complexes within host cell substructures. In this report, we describe experiments with coxsackie B viruses with a cell type-specific propagation deficit in Sk-N-Mc neuroblastoma cells conferred by the combination of a heterologous IRES and altered 3'UTR. Serial passage of these constructs in Sk-N-Mc cells yielded genetic adaptation by mutations within the viral nonstructural proteins 3A and 3C. Our data implicate 3A and/or 3C or their precursors 3AB and/or 3CD in a functional complex with the IRES and 3'UTR that drives viral propagation. Adaptation to neuroblastoma cells suggests an involvement of cell type-specific host factors or the host cell cytoplasmic milieu in this phenomenon.

Authors
Florez de Sessions, P; Dobrikova, E; Gromeier, M
MLA Citation
Florez de Sessions, P, Dobrikova, E, and Gromeier, M. "Genetic adaptation to untranslated region-mediated enterovirus growth deficits by mutations in the nonstructural proteins 3AB and 3CD." J Virol 81.16 (August 2007): 8396-8405.
PMID
17537861
Source
pubmed
Published In
Journal of virology
Volume
81
Issue
16
Publish Date
2007
Start Page
8396
End Page
8405
DOI
10.1128/JVI.00321-07

Viruses in the treatment of malignant glioma.

Authors
Everson, RG; Gromeier, M; Sampson, JH
MLA Citation
Everson, RG, Gromeier, M, and Sampson, JH. "Viruses in the treatment of malignant glioma." Expert Rev Neurother 7.4 (April 2007): 321-324. (Review)
PMID
17425484
Source
pubmed
Published In
Expert Review of Neurotherapeutics
Volume
7
Issue
4
Publish Date
2007
Start Page
321
End Page
324
DOI
10.1586/14737175.7.4.321

Attenuation of herpes simplex virus neurovirulence with picornavirus cis-acting genetic elements.

Viral pathogenesis depends on a suitable milieu in target host cells permitting viral gene expression, propagation, and spread. In many instances, viral genomes can be manipulated to select for propagation in certain tissues or cell types. This has been achieved for the neurotropic poliovirus (PV) by exchange of the internal ribosomal entry site (IRES), which is responsible for translation of the uncapped plus-strand RNA genome. The IRES of human rhinovirus type 2 (HRV2) confers neuron-specific replication deficits to PV but has no effect on viral propagation in malignant glioma cells. We report here that placing the critical gamma(1)34.5 virulence genes of herpes simplex virus type 1 (HSV) under translation control of the HRV2 IRES results in neuroattenuation in mice. In contrast, IRES insertion permits HSV propagation in malignant glioma cell lines that do not support replication of HSV recombinants carrying gamma(1)34.5 deletions. Our observations indicate that the conditions for alternative translation initiation at the HRV2 IRES in malignant glioma cells differ from those in normal central nervous system (CNS) cells. Picornavirus regulatory sequences mediating cell type-specific gene expression in the CNS can be utilized to target cancerous cells at the level of translation regulation outside their natural context.

Authors
Campbell, SA; Mulvey, M; Mohr, I; Gromeier, M
MLA Citation
Campbell, SA, Mulvey, M, Mohr, I, and Gromeier, M. "Attenuation of herpes simplex virus neurovirulence with picornavirus cis-acting genetic elements." J Virol 81.2 (January 2007): 791-799.
PMID
17079296
Source
pubmed
Published In
Journal of virology
Volume
81
Issue
2
Publish Date
2007
Start Page
791
End Page
799
DOI
10.1128/JVI.00714-06

The double-stranded RNA binding protein 76:NF45 heterodimer inhibits translation initiation at the rhinovirus type 2 internal ribosome entry site.

Poliovirus (PV) plus-strand RNA genomes initiate translation in a cap-independent manner via an internal ribosome entry site (IRES) in their 5' untranslated region. Viral translation is codetermined by cellular IRES trans-acting factors, which can influence viral propagation in a cell-type-specific manner. Engineering of a poliovirus recombinant devoid of neuropathogenic properties but highly lytic in malignant glioma cells was accomplished by exchange of the cognate poliovirus IRES with its counterpart from human rhinovirus type 2 (HRV2), generating PV-RIPO. Neuroblast:glioma heterokaryon analyses revealed that loss of neurovirulence is due to trans-dominant repression of PV-RIPO propagation in neuronal cells. The double-stranded RNA binding protein 76 (DRBP76) was previously identified to bind to the HRV2 IRES in neuronal cells and to inhibit PV-RIPO translation and propagation (M. Merrill, E. Dobrikova, and M. Gromeier, J. Virol. 80:3347-3356, 2006). The results of size exclusion chromatography indicate that DRBP76 heterodimerizes with nuclear factor of activated T cells, 45 kDa (NF45), in neuronal but not in glioma cells. The DRBP76:NF45 heterodimer binds to the HRV2 IRES in neuronal but not in glioma cells. Ribosomal profile analyses show that the heterodimer preferentially associates with the translation apparatus in neuronal cells and arrests translation at the HRV2 IRES, preventing PV-RIPO RNA assembly into polysomes. Results of this study suggest that the DRBP76:NF45 heterodimer selectively blocks HRV2 IRES-driven translation initiation in neuron-derived cells.

Authors
Merrill, MK; Gromeier, M
MLA Citation
Merrill, MK, and Gromeier, M. "The double-stranded RNA binding protein 76:NF45 heterodimer inhibits translation initiation at the rhinovirus type 2 internal ribosome entry site." J Virol 80.14 (July 2006): 6936-6942.
PMID
16809299
Source
pubmed
Published In
Journal of virology
Volume
80
Issue
14
Publish Date
2006
Start Page
6936
End Page
6942
DOI
10.1128/JVI.00243-06

Competitive translation efficiency at the picornavirus type 1 internal ribosome entry site facilitated by viral cis and trans factors.

Enteroviruses (EVs) overcome their host cells by usurping the translation machinery to benefit viral gene expression. This is accomplished through alternative translation initiation in a cap-independent manner at the viral internal ribosomal entry site (IRES). We have investigated the role of cis- and trans-acting viral factors in EV IRES translation in living cells. We observed that considerable portions of the viral genome, including the 5'-proximal open reading frame and the 3' untranslated region, contribute to stimulation of IRES-mediated translation. With the IRES in proper context, translation via internal initiation in uninfected cells is as efficient as at capped messages with short, unstructured 5' untranslated regions. IRES function is enhanced in cells infected with the EV coxsackievirus B3, but the related poliovirus has no significant stimulatory activity. This differential is due to the inherent properties of their 2A protease and is not coupled to 2A-mediated proteolytic degradation of the eukaryotic initiation factor 4G. Our results suggest that the efficiency of alternative translation initiation at EV IRESs depends on a properly configured template rather than on targeted alterations of the host cell translation machinery.

Authors
Dobrikova, EY; Grisham, RN; Kaiser, C; Lin, J; Gromeier, M
MLA Citation
Dobrikova, EY, Grisham, RN, Kaiser, C, Lin, J, and Gromeier, M. "Competitive translation efficiency at the picornavirus type 1 internal ribosome entry site facilitated by viral cis and trans factors." J Virol 80.7 (April 2006): 3310-3321.
PMID
16537598
Source
pubmed
Published In
Journal of virology
Volume
80
Issue
7
Publish Date
2006
Start Page
3310
End Page
3321
DOI
10.1128/JVI.80.7.3310-3321.2006

Cell-type-specific repression of internal ribosome entry site activity by double-stranded RNA-binding protein 76.

Translation of picornavirus plus-strand RNA genomes occurs via internal ribosomal entry at highly structured 5' untranslated regions. In addition to canonical translation factors, translation rate is likely influenced by supplementary host and viral trans-acting factors. We previously reported that insertion of a heterologous human rhinovirus type 2 internal ribosomal entry site (IRES) into the poliovirus (PV) genome, generating the chimeric virus PV-RIPO, selectively abrogates viral translation and propagation in neurons, which eliminate poliovirus's signature neuropathogenicity. While severely deficient in cells of neuronal lineage, the rhinovirus IRES promotes efficient propagation of PV-RIPO in cancer cells. Tumor-specific IRES function can be therapeutically exploited to direct viral cytotoxicity to cancer cells. Neuron-glioma heterokaryon analysis implicates neuronal trans-dominant inhibition in this effect, suggesting that host trans-acting factors repress IRES function in a cell-type-specific manner. We identified a set of proteins from neuronal cells with affinity for the rhinovirus IRES, including double-stranded RNA-binding protein 76 (DRBP76). DRBP76 associates with the IRES in neuronal but not in malignant glioma cells. Moreover, DRBP76 depletion in neuronal cells enhances rhinovirus IRES-driven translation and virus propagation. Our observations suggest that cell-type-specific association of DRBP76 with the rhinovirus IRES represses PV-RIPO translation and propagation in neuronal cells.

Authors
Merrill, MK; Dobrikova, EY; Gromeier, M
MLA Citation
Merrill, MK, Dobrikova, EY, and Gromeier, M. "Cell-type-specific repression of internal ribosome entry site activity by double-stranded RNA-binding protein 76." J Virol 80.7 (April 2006): 3147-3156.
PMID
16537583
Source
pubmed
Published In
Journal of virology
Volume
80
Issue
7
Publish Date
2006
Start Page
3147
End Page
3156
DOI
10.1128/JVI.80.7.3147-3156.2006

Targeted therapy for glioblastoma multiforme neoplastic meningitis with intrathecal delivery of an oncolytic recombinant poliovirus.

PURPOSE: The toxicity and antitumor activity of regional intrathecal delivery of an oncolytic recombinant poliovirus, PVS-RIPO, was evaluated in rodent models of glioblastoma multiforme neoplastic meningitis. EXPERIMENTAL DESIGN: To evaluate for toxicity, PVS-RIPO was administered into the spinal cord of transgenic mice that express the human poliovirus receptor, CD155, and into the intrathecal space of athymic rats without tumor. To evaluate efficacy, two different doses of PVS-RIPO were administered intrathecally 3 days after athymic rats were inoculated intrathecally with an aggressive human glioblastoma multiforme xenograft. RESULTS: No clinical or histologic evidence of toxicity was found. In efficacy studies, median survival was increased by 174.47% from 8.5 days in the group treated with UV light-inactivated virus to 15 days in the rats treated with 1.0 x 10(7) plaque-forming units (pfu) of PVS-RIPO (P < 0.0001). A similar increase in median survival was seen in the group receiving 1.0 x 10(9) pfu PVS-RIPO (P < 0.0001); however, there was no statistically significant dose-response relationship (P = 0.345). In addition, 1 of 10 rats in lower-dose PVS-RIPO-treated group and 3 of 10 rats in higher-dose PVS-RIPO-treated group survived >60 days after tumor cell inoculation and had no evidence of residual tumor at autopsy. CONCLUSION: These results suggest that intrathecal treatment with PVS-RIPO may be useful for treatment of neoplastic meningitis in patients with glioblastoma multiforme and provides a rationale for clinical trials in this area.

Authors
Ochiai, H; Campbell, SA; Archer, GE; Chewning, TA; Dragunsky, E; Ivanov, A; Gromeier, M; Sampson, JH
MLA Citation
Ochiai, H, Campbell, SA, Archer, GE, Chewning, TA, Dragunsky, E, Ivanov, A, Gromeier, M, and Sampson, JH. "Targeted therapy for glioblastoma multiforme neoplastic meningitis with intrathecal delivery of an oncolytic recombinant poliovirus." Clin Cancer Res 12.4 (February 15, 2006): 1349-1354.
PMID
16489093
Source
pubmed
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
12
Issue
4
Publish Date
2006
Start Page
1349
End Page
1354
DOI
10.1158/1078-0432.CCR-05-1595

The hepatitis C virus 3'-untranslated region or a poly(A) tract promote efficient translation subsequent to the initiation phase.

Enhancement of eukaryotic messenger RNA (mRNA) translation initiation by the 3' poly(A) tail is mediated through interaction of poly(A)-binding protein with eukaryotic initiation factor (eIF) 4G, bridging the 5' terminal cap structure. In contrast to cellular mRNA, translation of the uncapped, non-polyadenylated hepatitis C virus (HCV) genome occurs independently of eIF4G and a role for 3'-untranslated sequences in modifying HCV gene expression is controversial. Utilizing cell-based and in vitro translation assays, we show that the HCV 3'-untranslated region (UTR) or a 3' poly(A) tract of sufficient length interchangeably stimulate translation dependent upon the HCV internal ribosomal entry site (IRES). However, in contrast to cap-dependent translation, the rate of initiation at the HCV IRES was unaffected by 3'-untranslated sequences. Analysis of post-initiation events revealed that the 3' poly(A) tract and HCV 3'-UTR improve translation efficiency by enabling termination and possibly ribosome recycling for successive rounds of translation.

Authors
Bradrick, SS; Walters, RW; Gromeier, M
MLA Citation
Bradrick, SS, Walters, RW, and Gromeier, M. "The hepatitis C virus 3'-untranslated region or a poly(A) tract promote efficient translation subsequent to the initiation phase. (Published online)" Nucleic Acids Res 34.4 (2006): 1293-1303.
PMID
16510853
Source
pubmed
Published In
Nucleic Acids Research
Volume
34
Issue
4
Publish Date
2006
Start Page
1293
End Page
1303
DOI
10.1093/nar/gkl019

Oncolytic viruses for the treatment of malignant glioma

Malignant glioma is the most common primary malignancy of the human CNS. Despite decades of research, the current therapeutic strategy consists of a multimodal regimen of surgery, chemotherapy and radiation. This course of therapy yields a median survival after diagnosis of ∼ 1 year. This dismal prognosis inspires the ongoing development of novel oncolytic agents targeting glioma, which include gene therapy, immunomodulatory therapy and oncolytic viruses. Oncolytic viruses are defined by their ability to target, replicate in and lyse tumour cells without critically damaging surrounding non-cancerous tissue. Although some viruses are naturally oncolytic and tumour-selective, the advent of modern recombinant DNA technology has allowed the engineering of additional viruses with improved therapeutic indices. This technological advance has enabled rapid growth in the field of viral therapy. Reovirus, Newcastle disease virus (NDV), measles virus, adenovirus, poliovirus and herpes simplex virus-1 are in preclinical and clinical development for use as oncolytic agents against malignant glioma. This report will focus on the recent patent literature in the field of oncolytic viruses for the treatment of malignant glioma. © 2006 Ashley Publications.

Authors
Merrill, MK; Selznick, LA; Gromeier, M
MLA Citation
Merrill, MK, Selznick, LA, and Gromeier, M. "Oncolytic viruses for the treatment of malignant glioma." Expert Opinion on Therapeutic Patents 16.3 (2006): 363-371.
Source
scival
Published In
Expert Opinion on Therapeutic Patents
Volume
16
Issue
3
Publish Date
2006
Start Page
363
End Page
371
DOI
10.1517/13543776.16.3.363

Understanding polio: New insights from a cold virus

Authors
Dufresne, AT; Gromeier, M
MLA Citation
Dufresne, AT, and Gromeier, M. "Understanding polio: New insights from a cold virus." Microbe 1.1 (2006): 13-18.
Source
scival
Published In
Microbe (Washington, D.C.)
Volume
1
Issue
1
Publish Date
2006
Start Page
13
End Page
18

The polypyrimidine tract binding protein is required for efficient picornavirus gene expression and propagation.

Mammalian host factors required for efficient viral gene expression and propagation have been often recalcitrant to genetic analysis. A case in point is the function of cellular factors that trans-activate internal ribosomal entry site (IRES)-driven translation, which is operative in many positive-stranded RNA viruses, including all picornaviruses. These IRES trans-acting factors have been elegantly studied in vitro, but their in vivo importance for viral gene expression and propagation has not been widely confirmed experimentally. Here we use RNA interference to deplete mammalian cells of one such factor, the polypyrimidine tract binding protein, and test its requirement in picornavirus gene expression and propagation. Depletion of the polypyrimidine tract binding protein resulted in a marked delay of particle propagation and significantly decreased synthesis and accumulation of viral proteins of poliovirus and encephalomyocarditis virus. These effects could be partially restored by expression of an RNA interference-resistant exogenous polypyrimidine tract binding protein. These data indicate a critical role for the polypyrimidine tract binding protein in picornavirus gene expression and strongly suggest a requirement for efficient IRES-dependent translation.

Authors
Florez, PM; Sessions, OM; Wagner, EJ; Gromeier, M; Garcia-Blanco, MA
MLA Citation
Florez, PM, Sessions, OM, Wagner, EJ, Gromeier, M, and Garcia-Blanco, MA. "The polypyrimidine tract binding protein is required for efficient picornavirus gene expression and propagation." J Virol 79.10 (May 2005): 6172-6179.
PMID
15858002
Source
pubmed
Published In
Journal of virology
Volume
79
Issue
10
Publish Date
2005
Start Page
6172
End Page
6179
DOI
10.1128/JVI.79.10.6172-6179.2005

Genetic determinants of cell type-specific poliovirus propagation in HEK 293 cells.

The ability of poliovirus to propagate in neuronal cells can be reduced by introducing appropriate nucleotide substitutions into the viral genome. Specific mutations scattered throughout the poliovirus genome yielded the live attenuated vaccine strains of poliovirus. Neuron-specific propagation deficits of the Sabin strains are partially encrypted within a confined region of the internal ribosomal entry site (IRES), which carries attenuating point mutations in all three serotypes. Recently, high levels of neurovirulence attenuation were achieved with genetically engineered polioviruses containing heterologous IRES elements. This is exemplified with poliovirus recombinants replicating under control of a human rhinovirus type 2 (HRV2) IRES element. We have carried out experiments delineating the genetic basis for neuronal IRES function. Neuronal dysfunction of the HRV2 IRES is determined mainly by IRES stem-loop domain V, the locus for attenuating point mutations within the Sabin strains. Neuronal incompetence associated with HRV2 IRES domain V is substantially more pronounced than that observed with the attenuating IRES point mutation of the Sabin serotype 1 vaccine strain. Mix-and-match recombination of polio and HRV2 IRES domain V suggests that the attenuation phenotype correlates with overall structural features rather than primary sequence. Our experiments have identified HEK 293 cells as a novel system for the study of neuron-specific replication phenotypes of poliovirus. This cell line, originally derived from embryonic human kidney, has recently been described to display neuronal characteristics. We report propagation properties in HEK 293 cells for poliovirus recombinants with attenuated neurovirulence in experimental animals that corroborate this observation.

Authors
Campbell, SA; Lin, J; Dobrikova, EY; Gromeier, M
MLA Citation
Campbell, SA, Lin, J, Dobrikova, EY, and Gromeier, M. "Genetic determinants of cell type-specific poliovirus propagation in HEK 293 cells." J Virol 79.10 (May 2005): 6281-6290.
PMID
15858012
Source
pubmed
Published In
Journal of virology
Volume
79
Issue
10
Publish Date
2005
Start Page
6281
End Page
6290
DOI
10.1128/JVI.79.10.6281-6290.2005

Oncolytic viruses for cancer therapy II. Cell-internal factors for conditional growth in neoplastic cells.

Recent advances in our understanding of virus-host interactions have fueled new studies in the field of oncolytic viruses. The first part of this review explained how cell-external factors, such as cellular receptors, influence tumor tropism and specificity of oncolytic virus candidates. In the second part of this review, we focus on cellinternal factors that mediate tumor-specific virus growth. An oncolytic virus must be able to replicate within cancerous cells and kill them without collateral damage to healthy surrounding cells. This desirable property is inherent to some proposed oncolytic viral agents or has been achieved by genetic manipulation in others.

Authors
Campbell, SA; Gromeier, M
MLA Citation
Campbell, SA, and Gromeier, M. "Oncolytic viruses for cancer therapy II. Cell-internal factors for conditional growth in neoplastic cells." Onkologie 28.4 (April 2005): 209-215. (Review)
PMID
15840970
Source
pubmed
Published In
Onkologie
Volume
28
Issue
4
Publish Date
2005
Start Page
209
End Page
215
DOI
10.1159/000084010

Oncolytic viruses for cancer therapy I. Cell-external factors: virus entry and receptor interaction.

After being recognized for their anti-neoplastic properties at the beginning of the last century, viruses are again being considered for use as therapeutic agents against cancer. Certain virus species have a propensity to replicate within transformed cells, which are commonly rendered vulnerable due to tumor-specific defects in their defense against viral infection. Other viruses have been modified to subject them to tumor-specific growth conditions. Oncolytic viruses carry the promise to efficiently target cancer cells for destruction and spread throughout tumor tissue to reach distant neoplastic loci without causing collateral damage to healthy tissues. In contrast to conventional cancer chemotherapy, viral anti-neoplastic agents require complex interactions with the host organism to reach their target and to unfold their oncolytic activity. Recent progress in the elucidation of the molecular mechanisms of viral pathogenesis has opened up new opportunities to manipulate virus-host interactions, generating effective anti-tumor strategies. On the other hand, significant obstacles towards the application of safe and efficacious viral therapies have become apparent. These frequently relate to the lack of cell culture and animal tumor models that accurately reflect the characteristics of cancerous tissues in patients. Throughout the past century, viral therapeutics against cancer have evolved into a new class of treatment strategies characterized by unique opportunities and challenges. A growing number of oncolytic viruses has entered clinical investigation or is scheduled to do so in the near future. Great efforts are being undertaken to rekindle an old idea and, with the help of new technologies, to realize its promise of new treatment facilities for cancer.

Authors
Campbell, SA; Gromeier, M
MLA Citation
Campbell, SA, and Gromeier, M. "Oncolytic viruses for cancer therapy I. Cell-external factors: virus entry and receptor interaction." Onkologie 28.3 (March 2005): 144-149. (Review)
PMID
15772465
Source
pubmed
Published In
Onkologie
Volume
28
Issue
3
Publish Date
2005
Start Page
144
End Page
149
DOI
10.1159/000083659

Treatment of CNS malignancies with a recombinant oncolytic poliovirus

Spread to the central nervous system (CNS) and the leptomeninges is a frequent complication of systemic cancers that is associated with serious morbidity and high mortality. We have developed a novel therapeutic approach against primary CNS tumors and secondary CNS malignancy complicating systemic cancer based on the human neuropathogen poliovirus. Susceptibility to poliovirus infection and ensuing rapid cell lysis are mediated by the cellular poliovirus receptor CD155, as well as cell-internal factors involved in control of viral gene expression and genome replication. We observed that CD155, a putative cell adhesion molecule of the immunoglobulin superfamily physiologically expressed in the developing CNS, is ectopically expressed on numerous primary and secondary CNS neoplasms. An association with malignant cells renders CD155 a molecular target for therapeutic intervention with poliovirus. However, any therapeutic application of poliovirus must contend with its inherent neuropathogenic potential. Poliovirus gene expression is driven by the viral internal ribosomal entry site (IRES), which mediates translation initiation in a 5'-end, cap-independent manner. We discovered that IRES function is subject to potent cell type-specific restrictions that can be exploited to selectively drive viral gene expression and propagation in cancerous cells. Polioviruses replicating under control of a heterologous IRES derived from their relative human rhinovirus type 2, displayed rapid growth and cell killing in cancerous cells. However, these recombinants failed to efficiently translate their genome and propagate in cells of neuronal lineage. Consequently, polio-/rhinovirus recombinants are non-pathogenic after intraspinal inoculation into mice transgenic for the poliovirus receptor CD155 and in non-human primates. Oncolytic poliovirus recombinants have shown promise in preclinical investigations against primary tumor explant cultures and in xenograft animal studies and are currently being prepared for phase-I clinical trials against glioblastoma multiforme. © 2005 Bentham Science Publishers Ltd.

Authors
Moore, SA; Merrill, MK; Gromeier, M
MLA Citation
Moore, SA, Merrill, MK, and Gromeier, M. "Treatment of CNS malignancies with a recombinant oncolytic poliovirus." Drug Design Reviews Online 2.3 (2005): 233-241.
Source
scival
Published In
Drug Design Reviews Online
Volume
2
Issue
3
Publish Date
2005
Start Page
233
End Page
241
DOI
10.2174/1567269053828774

A nonpolio enterovirus with respiratory tropism causes poliomyelitis in intercellular adhesion molecule 1 transgenic mice.

Coxsackievirus A21 (CAV21) is classified within the species Human enterovirus C (HEV-C) of the Enterovirus genus of picornaviruses. HEV-C share striking homology with the polioviruses (PV), their closest kin among the enteroviruses. Despite a high level of sequence identity, CAV21 and PV cause distinct clinical disease typically attributed to their differential use of host receptors. PV cause poliomyelitis, whereas CAV21 shares a receptor and a propensity to cause upper respiratory tract infections with the major group rhinoviruses. As a model for CAV21 infection, we have developed transgenic mice that express human intercellular adhesion molecule 1, the cell-surface receptor for CAV21. Surprisingly, CAV21 administered to these mice via the intramuscular route causes a paralytic condition consistent with poliomyelitis. The virus appears to invade the CNS by retrograde axonal transport, as has been demonstrated to occur in analogous PV infections. We detected human intercellular adhesion molecule 1 expression on both transgenic mouse and human spinal cord anterior horn motor neurons, indicating that members of HEV-C may share PV's potential to elicit poliomyelitis in humans.

Authors
Dufresne, AT; Gromeier, M
MLA Citation
Dufresne, AT, and Gromeier, M. "A nonpolio enterovirus with respiratory tropism causes poliomyelitis in intercellular adhesion molecule 1 transgenic mice." Proc Natl Acad Sci U S A 101.37 (September 14, 2004): 13636-13641.
PMID
15353596
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
101
Issue
37
Publish Date
2004
Start Page
13636
End Page
13641
DOI
10.1073/pnas.0403998101

Treatment of intracerebral neoplasia and neoplastic meningitis with regional delivery of oncolytic recombinant poliovirus.

PURPOSE: Spread to the central nervous system (CNS) and the leptomeninges is a frequent complication of systemic cancers that is associated with serious morbidity and high mortality. We have evaluated a novel therapeutic approach against CNS complications of breast cancer based on the human neuropathogen poliovirus (PV). EXPERIMENTAL DESIGN: Susceptibility to PV infection and ensuing rapid cell lysis is mediated by the cellular receptor of PV, CD155. We evaluated CD155 expression in several human breast tumor tissue specimens and cultured breast cancer cell lines. In addition, we tested an oncolytic PV recombinant for efficacy in xenotransplantation models of neoplastic meningitis and cerebral metastasis secondary to breast cancer. RESULTS: We observed that breast cancer tissues and cell lines derived thereof express CD155 at levels mediating exquisite sensitivity toward PV-induced oncolysis in the latter. An association with the immunoglobulin superfamily molecule CD155 renders breast cancer a likely target for oncolytic PV recombinants. This assumption was confirmed in xenotransplantation models for neoplastic meningitis or solitary cerebral metastasis, where local virus treatment dramatically improved survival. CONCLUSIONS: Our findings suggest oncolytic PV recombinants as a viable treatment option for CNS complications of breast cancer.

Authors
Ochiai, H; Moore, SA; Archer, GE; Okamura, T; Chewning, TA; Marks, JR; Sampson, JH; Gromeier, M
MLA Citation
Ochiai, H, Moore, SA, Archer, GE, Okamura, T, Chewning, TA, Marks, JR, Sampson, JH, and Gromeier, M. "Treatment of intracerebral neoplasia and neoplastic meningitis with regional delivery of oncolytic recombinant poliovirus." Clin Cancer Res 10.14 (July 15, 2004): 4831-4838.
PMID
15269159
Source
pubmed
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
10
Issue
14
Publish Date
2004
Start Page
4831
End Page
4838
DOI
10.1158/1078-0432.CCR-03-0694

Poliovirus receptor CD155-targeted oncolysis of glioma.

Cell adhesion molecules of the immunoglobulin superfamily are aberrantly expressed in malignant glioma. Amongst these, the human poliovirus receptor CD155 provides a molecular target for therapeutic intervention with oncolytic poliovirus recombinants. Poliovirus has been genetically modified through insertion of regulatory sequences derived from human rhinovirus type 2 to selectively replicate within and destroy cancerous cells. Efficacious oncolysis mediated by poliovirus derivatives depends on the presence of CD155 in targeted tumors. To prepare oncolytic polioviruses for clinical application, we have developed a series of assays in high-grade malignant glioma (HGL) to characterize CD155 expression levels and susceptibility to oncolytic poliovirus recombinants. Analysis of 6 HGL cases indicates that CD155 is expressed in these tumors and in primary cell lines derived from these tumors. Upregulation of the molecular target CD155 rendered explant cultures of all studied tumors highly susceptible to a prototype oncolytic poliovirus recombinant. Our observations support the clinical application of such agents against HGL.

Authors
Merrill, MK; Bernhardt, G; Sampson, JH; Wikstrand, CJ; Bigner, DD; Gromeier, M
MLA Citation
Merrill, MK, Bernhardt, G, Sampson, JH, Wikstrand, CJ, Bigner, DD, and Gromeier, M. "Poliovirus receptor CD155-targeted oncolysis of glioma." Neuro Oncol 6.3 (July 2004): 208-217.
PMID
15279713
Source
pubmed
Published In
Neuro-Oncology
Volume
6
Issue
3
Publish Date
2004
Start Page
208
End Page
217
DOI
10.1215/S1152851703000577

The double-stranded RNA-activated protein kinase mediates viral-induced encephalitis.

The double-stranded (ds) RNA-activated protein kinase (PKR) plays an important role in control of viral infections and cell growth. We have studied the role of PKR in viral infection in mice that are defective in the PKR signaling pathway. Transgenic mice were derived that constitutively express a trans-dominant-negative kinase-defective mutant PKR under control of the beta-actin promoter. The trans-dominant-negative PKR mutant expressing transgenic mice do not have a detectable phenotype, similar to observations with PKR knock-out mice. The requirement for PKR in viral pathogenesis was studied by intracerebral infection of mice with a mouse-adapted poliovirus. Histopathological analysis revealed diffuse encephalomyelitis with severe inflammatory lesions throughout the central nervous system (CNS) in infected wild-type mice. In contrast, histopathological evaluation of virus-injected trans-dominant-negative PKR transgenic mice as well as PKR knock-out mice yielded no signs of tissue damage associated with inflammatory host responses. However, the virus did replicate in both models of PKR-deficient mice at a level equal to that observed in wild-type infected mice. Although the results indicate a clear difference in susceptibility to poliovirus-induced encephalitis, this difference manifests clinically as a slight delay in fatal neuropathy in trans-dominant-negative PKR transgenic and PKR knock-out animals. Our observations support the finding that viral-induced PKR activation may play a significant role in pathogenesis by mediating the host response to viral CNS infection. They support PKR to be an effective target to control tissue damage due to deleterious host responses to viral infection.

Authors
Scheuner, D; Gromeier, M; Davies, MV; Dorner, AJ; Song, B; Patel, RV; Wimmer, EJ; McLendon, RE; Kaufman, RJ
MLA Citation
Scheuner, D, Gromeier, M, Davies, MV, Dorner, AJ, Song, B, Patel, RV, Wimmer, EJ, McLendon, RE, and Kaufman, RJ. "The double-stranded RNA-activated protein kinase mediates viral-induced encephalitis." Virology 317.2 (December 20, 2003): 263-274.
PMID
14698665
Source
pubmed
Published In
Virology
Volume
317
Issue
2
Publish Date
2003
Start Page
263
End Page
274

Activity of a type 1 picornavirus internal ribosomal entry site is determined by sequences within the 3' nontranslated region.

We have proposed a cancer treatment modality based on poliovirus chimeras replicating under the translational control of an internal ribosomal entry site (IRES) derived from human rhinovirus type 2. Insertion of the heterologous IRES causes a neuron-specific propagation deficit and eliminates neurovirulence inherent in poliovirus without affecting viral growth in cells derived from malignant gliomas. We now report the elucidation of a molecular mechanism responsible for the cell type-specific defect mediated by the rhinovirus IRES. Rhinovirus IRES function in neuronal cell types depends on specific structural elements within the 3' non-translated region of the viral genome. Our observations suggest long-range interactions between the IRES and the 3' terminus that control IRES-mediated gene expression and virus propagation.

Authors
Dobrikova, E; Florez, P; Bradrick, S; Gromeier, M
MLA Citation
Dobrikova, E, Florez, P, Bradrick, S, and Gromeier, M. "Activity of a type 1 picornavirus internal ribosomal entry site is determined by sequences within the 3' nontranslated region." Proc Natl Acad Sci U S A 100.25 (December 9, 2003): 15125-15130.
PMID
14645707
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
100
Issue
25
Publish Date
2003
Start Page
15125
End Page
15130
DOI
10.1073/pnas.2436464100

Structural determinants of insert retention of poliovirus expression vectors with recombinant IRES elements.

Although picornaviruses provide attractive vectors for expression of foreign genes, poor genetic stability restricts their use for immunization purposes. A new prototype vector was generated to increase foreign insert retention, by shifting of the initiation codon to a cryptic AUG within the internal ribosomal entry site (IRES) and replacement of IRES domain VI with foreign ORFs. Using our strategy to replace regulatory noncoding sequences with unrelated foreign genetic material, we generated stable poliovirus-based expression vectors with robust long-term expression of foreign ORFs. Our studies revealed that size and predicted secondary structure formed by the heterologous sequences govern long-term retention and efficiency of expression of foreign inserts replacing IRES structures. These observations indicate that, with certain limitations imposed by structural preferences, foreign sequences can functionally replace IRES substructures in stable picornavirus immunization vectors.

Authors
Dobrikova, EY; Florez, P; Gromeier, M
MLA Citation
Dobrikova, EY, Florez, P, and Gromeier, M. "Structural determinants of insert retention of poliovirus expression vectors with recombinant IRES elements." Virology 311.2 (July 5, 2003): 241-253.
PMID
12842615
Source
pubmed
Published In
Virology
Volume
311
Issue
2
Publish Date
2003
Start Page
241
End Page
253

Oncolytic Viruses for Cancer Therapy

After being recognized for their antineoplastic properties at the beginning of the last century, viruses are again being considered for use as therapeutic agents against cancer. Certain species of virus have a propensity to replicate within transformed cells, commonly rendered vulnerable because of tumor-specific defects in their defense against viral infection. Other viruses have been modified to tumor-specific growth conditions. Oncolytic viruses carry the promise to efficiently target cancer cells for destruction and spread throughout tumor tissue to reach distant neoplastic loci without causing collateral damage to healthy tissues. In contrast to conventional cancer chemotherapy, viral antineoplastic agents require complex interactions with the host organism to reach their target and to develop oncolytic activity. Recent progress in the elucidation of the molecular mechanisms of viral pathogenesis has opened up new opportunities to manipulate virus-host interactions, generating effective antitumor strategies. On the other hand, significant obstacles towards the application of safe and efficacious viral therapies have become apparent. These frequently relate to the lack of cell culture and animal tumor models that accurately reflect the characteristics of cancerous tissues in patients. Throughout the past century, viral therapeutics against cancer has evolved into a new class of treatment strategies characterized by unique opportunities and challenges. A growing number of oncolytic viruses have entered clinical investigation or are scheduled to do so in the near future. Great efforts are being undertaken to rekindle an old idea and, with the help of new technologies, to realize its promise of new treatment options for cancer.

Authors
Gromeier, M
MLA Citation
Gromeier, M. "Oncolytic Viruses for Cancer Therapy." American Journal of Cancer 2.5 (2003): 313-323.
Source
scival
Published In
American Journal of Cancer
Volume
2
Issue
5
Publish Date
2003
Start Page
313
End Page
323
DOI
10.2165/00024669-200302050-00002

Viruses in the treatment of brain tumors.

The grave outlook for malignant glioma patients in spite of improvements to current modalities has ushered in new approaches to therapy. Viruses have emerged on the scene and gained attention for their ability to play essentially two roles: first, as vectors for therapeutic gene delivery and second, as engineered infectious agents capable of selectively lysing tumor cells. To date, clinical brain tumor trials using viruses for gene delivery have employed retroviral or adenoviral vectors to introduce ganciclovir susceptibility to tumors in the form of the HSV1-TK gene. Clinical oncolytic studies, on the other hand, have evaluated a conditionally replicating HSV as an antineoplastic agent. Despite some promise afforded by these trials, further studies are warranted; the investigation of additional viruses to play these roles is inevitable and is now precedented.

Authors
Fecci, PE; Gromeier, M; Sampson, JH
MLA Citation
Fecci, PE, Gromeier, M, and Sampson, JH. "Viruses in the treatment of brain tumors." Neuroimaging Clin N Am 12.4 (November 2002): 553-570. (Review)
PMID
12687911
Source
pubmed
Published In
Neuroimaging Clinics of North America
Volume
12
Issue
4
Publish Date
2002
Start Page
553
End Page
570

Genetically stable picornavirus expression vectors with recombinant internal ribosomal entry sites.

In many respects, picornaviruses are well suited for their proposed use as immunization vectors. However, their inherent genetic instability hinders application for prophylactic purposes. We demonstrate the improved expression and stability of a heterologous insert through a novel vector design strategy that partially replaces noncoding regulatory sequences with coding sequences for foreign gene products.

Authors
Dufresne, AT; Dobrikova, EY; Schmidt, S; Gromeier, M
MLA Citation
Dufresne, AT, Dobrikova, EY, Schmidt, S, and Gromeier, M. "Genetically stable picornavirus expression vectors with recombinant internal ribosomal entry sites." J Virol 76.17 (September 2002): 8966-8972.
PMID
12163617
Source
pubmed
Published In
Journal of virology
Volume
76
Issue
17
Publish Date
2002
Start Page
8966
End Page
8972

Expression of the human poliovirus receptor/CD155 gene is activated by sonic hedgehog.

The human poliovirus receptor/CD155 is a transmembrane glycoprotein belonging to the immunoglobulin superfamily. The ectodomain of CD155 mediates cell attachment to the extracellular matrix molecule vitronectin, while its intracellular domain interacts with the dynein light chain Tctex-1. CD155 is a primate-restricted gene that is expressed during development in mesenchymal tissues and ventrally derived structures within the CNS. Its function in adults is as yet unknown, but significantly, CD155 is aberrantly expressed in neuroectodermal tumors. We show that the expression of CD155 mRNA is up-regulated when human Ntera2 cells are treated with purified Sonic hedgehog (Shh) protein. Reporter gene expression driven by the CD155 core promoter is activated by Shh in transient co-transfection assays. Analysis of the CD155 core promoter indicates that an intact GLI binding site is required for Shh activation. In addition, overexpression of Gli1 or Gli3 potently activates reporter gene expression driven by the CD155 core promoter. These data identify the CD155 gene as a transcriptional target of Shh, a finding that has significance for the normal function of CD155 during development and the expression of CD155 in neuroectodermal tumors.

Authors
Solecki, DJ; Gromeier, M; Mueller, S; Bernhardt, G; Wimmer, E
MLA Citation
Solecki, DJ, Gromeier, M, Mueller, S, Bernhardt, G, and Wimmer, E. "Expression of the human poliovirus receptor/CD155 gene is activated by sonic hedgehog." J Biol Chem 277.28 (July 12, 2002): 25697-25702.
PMID
11983699
Source
pubmed
Published In
The Journal of biological chemistry
Volume
277
Issue
28
Publish Date
2002
Start Page
25697
End Page
25702
DOI
10.1074/jbc.M201378200

Viruses for the treatment of malignant glioma.

Viruses have been considered for use as therapeutic agents against cancer, and malignant glioma in particular. Oncolytic viruses were designed to target malignant cells supporting efficient virus replication, or rendered vulnerable to viral destruction due to tumor-specific defects in their defense against virus infection. Other than conventional cancer chemotherapy, viral antineoplastic agents require complex interactions with the host organism to reach their target and to unleash their oncolytic activity. Recent progress in the design and therapeutic application of oncolytic viruses carries the promise to make these agents available for treatment of malignant glioma.

Authors
Gromeier, M; Wimmer, E
MLA Citation
Gromeier, M, and Wimmer, E. "Viruses for the treatment of malignant glioma." Curr Opin Mol Ther 3.5 (October 2001): 503-508. (Review)
PMID
11699896
Source
pubmed
Published In
Current Opinion in Molecular Therapeutics
Volume
3
Issue
5
Publish Date
2001
Start Page
503
End Page
508

Viruses as therapeutic agents against malignant disease of the central nervous system.

Authors
Gromeier, M
MLA Citation
Gromeier, M. "Viruses as therapeutic agents against malignant disease of the central nervous system." J Natl Cancer Inst 93.12 (June 20, 2001): 889-890.
PMID
11416101
Source
pubmed
Published In
Journal of the National Cancer Institute
Volume
93
Issue
12
Publish Date
2001
Start Page
889
End Page
890

Inactivated vaccines based on alternatives to wild-type seed virus.

Authors
Chumakov, K; Dragunsky, E; Ivshina, A; Enterline, J; Wells, V; Nomura, T; Gromeier, M; Wimmer, E
MLA Citation
Chumakov, K, Dragunsky, E, Ivshina, A, Enterline, J, Wells, V, Nomura, T, Gromeier, M, and Wimmer, E. "Inactivated vaccines based on alternatives to wild-type seed virus." Dev Biol (Basel) 105 (2001): 171-177.
PMID
11763325
Source
pubmed
Published In
Developments in biologicals
Volume
105
Publish Date
2001
Start Page
171
End Page
177

Expression of the human poliovirus receptor/CD155 gene during development of the central nervous system: implications for the pathogenesis of poliomyelitis.

The gene for the human poliovirus receptor (hPVR/CD155) is the founding member of a new family of genes encoding proteins belonging to the immunoglobulin superfamily. To determine whether CD155 is expressed during mammalian development, we have made use of the previously characterized promoter of the CD155 gene and generated mice transgenic for a CD155 promoter-driven beta-galactosidase reporter gene. Expression of the reporter gene in transgenic embryos was observed during midgestation in anterior midline structures of the developing central nervous system and in the neuroretina. During that period, reporter gene expression appeared within the notochord and floor plate along the entire spinal cord reaching into the caudal diencephalon. In addition, transgene expression was observed in axonal projections emanating from retinal ganglion cells forming the optic nerve to reach the future region of the optic chiasm. Analysis of expression of CD155 during human embryonic development confirmed the distribution of reporter gene expression specified by CD155 promoter activity. The anatomical distribution of CD155 promoter activity during embryogenesis matches that of transacting factors previously identified to regulate transcription of the CD155 gene. Expression of CD155 within embryonic structures giving rise to spinal cord anterior horn motor neurons may explain the restrictive host cell tropism of poliovirus for this cellular compartment of the CNS.

Authors
Gromeier, M; Solecki, D; Patel, DD; Wimmer, E
MLA Citation
Gromeier, M, Solecki, D, Patel, DD, and Wimmer, E. "Expression of the human poliovirus receptor/CD155 gene during development of the central nervous system: implications for the pathogenesis of poliomyelitis." Virology 273.2 (August 1, 2000): 248-257.
PMID
10915595
Source
pubmed
Published In
Virology
Volume
273
Issue
2
Publish Date
2000
Start Page
248
End Page
257
DOI
10.1006/viro.2000.0418

Intergeneric poliovirus recombinants for the treatment of malignant glioma.

Poliovirus neuropathogenicity depends on sequences within the 5' nontranslated region of the virus. Exchange of the poliovirus internal ribosomal entry site with its counterpart from human rhinovirus type 2 resulted in attenuation of neurovirulence in primates. Despite deficient virus propagation in cells of neuronal origin, nonpathogenic polio recombinants retain excellent growth characteristics in cell lines derived from glial neoplasms. Susceptibility of malignant glioma cells to poliovirus may be mediated by expression of a poliovirus receptor, CD155, in glial neoplasms. Intergeneric polio recombinants with heterologous internal ribosomal entry site elements unfolded strong oncolytic potential against experimentally induced gliomas in athymic mice. Our observations suggest that highly attenuated poliovirus recombinants may have applicability as biotherapeutic antineoplastic agents.

Authors
Gromeier, M; Lachmann, S; Rosenfeld, MR; Gutin, PH; Wimmer, E
MLA Citation
Gromeier, M, Lachmann, S, Rosenfeld, MR, Gutin, PH, and Wimmer, E. "Intergeneric poliovirus recombinants for the treatment of malignant glioma." Proc Natl Acad Sci U S A 97.12 (June 6, 2000): 6803-6808.
PMID
10841575
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
97
Issue
12
Publish Date
2000
Start Page
6803
End Page
6808

Dual stem loops within the poliovirus internal ribosomal entry site control neurovirulence.

In the human central nervous system, susceptibility to poliovirus (PV) infection is largely confined to a specific subpopulation of neuronal cells. PV tropism is likely to be determined by cell-external components such as the PV receptor CD155, as well as cell-internal constraints such as the availability of a suitable microenvironment for virus propagation. We reported previously that the exchange of the cognate internal ribosomal entry site (IRES) within the 5' nontranslated region of PV with its counterpart from human rhinovirus type 2 (HRV2) can eliminate the neuropathogenic phenotype in a transgenic mouse model for poliomyelitis without diminishing the growth properties in HeLa cells. We now show that attenuation of neurovirulence of PV/HRV2 chimeras is not confined to CD155 transgenic mice but is evident also after intraspinal inoculation into Cynomolgus monkeys. We have dissected the PV and HRV2 IRES elements to determine those structures responsible for neurovirulence (or attenuation) of these chimeric viruses. We report that two adjacent stem loop structures within the IRES cooperatively determine neuropathogenicity.

Authors
Gromeier, M; Bossert, B; Arita, M; Nomoto, A; Wimmer, E
MLA Citation
Gromeier, M, Bossert, B, Arita, M, Nomoto, A, and Wimmer, E. "Dual stem loops within the poliovirus internal ribosomal entry site control neurovirulence." J Virol 73.2 (February 1999): 958-964.
PMID
9882296
Source
pubmed
Published In
Journal of virology
Volume
73
Issue
2
Publish Date
1999
Start Page
958
End Page
964

The relation of prophylactic inoculations to the onset of poliomyelitis

Authors
McCloskey, BP; Gromeier, M; Wimmer, E
MLA Citation
McCloskey, BP, Gromeier, M, and Wimmer, E. "The relation of prophylactic inoculations to the onset of poliomyelitis." Reviews in Medical Virology 9.4 (1999): 219-226.
PMID
10578117
Source
scival
Published In
Reviews in Medical Virology
Volume
9
Issue
4
Publish Date
1999
Start Page
219
End Page
226
DOI
10.1002/(SICI)1099-1654(199910/12)9:4<219::AID-RMV249>3.0.CO;2-T

A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element.

Hepatitis C virus (HCV) infection frequently leads to chronic hepatitis and cirrhosis of the liver and has been linked to development of hepatocellular carcinoma. We previously identified a small yeast RNA (IRNA) capable of specifically inhibiting poliovirus (PV) internal ribosome entry site (IRES)-mediated translation. Here we report that IRNA specifically inhibits HCV IRES-mediated translation both in vivo and in vitro. A number of human hepatoma (Huh-7) cell lines expressing IRNA were prepared and characterized. Constitutive expression of IRNA was not detrimental to cell growth. HCV IRES-mediated cap-independent translation was markedly inhibited in cells constitutively expressing IRNA compared to control hepatoma cells. However, cap-dependent translation was not significantly affected in these cell lines. Additionally, Huh-7 cells constitutively expressing IRNA became refractory to infection by a PV-HCV chimera in which the PV IRES is replaced by the HCV IRES. In contrast, replication of a PV-encephalomyocarditis virus (EMCV) chimera containing the EMCV IRES element was not affected significantly in the IRNA-producing cell line. Finally, the binding of the La autoantigen to the HCV IRES element was specifically and efficiently competed by IRNA. These results provide a basis for development of novel drugs effective against HCV infection.

Authors
Das, S; Ott, M; Yamane, A; Tsai, W; Gromeier, M; Lahser, F; Gupta, S; Dasgupta, A
MLA Citation
Das, S, Ott, M, Yamane, A, Tsai, W, Gromeier, M, Lahser, F, Gupta, S, and Dasgupta, A. "A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element." J Virol 72.7 (July 1998): 5638-5647.
PMID
9621022
Source
pubmed
Published In
Journal of virology
Volume
72
Issue
7
Publish Date
1998
Start Page
5638
End Page
5647

Mechanism of injury-provoked poliomyelitis.

Skeletal muscle injury is known to predispose its sufferers to neurological complications of concurrent poliovirus infections. This phenomenon, labeled "provocation poliomyelitis," continues to cause numerous cases of childhood paralysis due to the administration of unnecessary injections to children in areas where poliovirus is endemic. Recently, it has been reported that intramuscular injections may also increase the likelihood of vaccine-associated paralytic poliomyelitis in recipients of live attenuated poliovirus vaccines. We have studied this important risk factor for paralytic polio in an animal system for poliomyelitis and have determined the pathogenic mechanism linking intramuscular injections and provocation poliomyelitis. Skeletal muscle injury induces retrograde axonal transport of poliovirus and thereby facilitates viral invasion of the central nervous system and the progression of spinal cord damage. The pathogenic mechanism of provocation poliomyelitis may differ from that of polio acquired in the absence of predisposing factors.

Authors
Gromeier, M; Wimmer, E
MLA Citation
Gromeier, M, and Wimmer, E. "Mechanism of injury-provoked poliomyelitis." J Virol 72.6 (June 1998): 5056-5060.
PMID
9573275
Source
pubmed
Published In
Journal of virology
Volume
72
Issue
6
Publish Date
1998
Start Page
5056
End Page
5060

Poliovirus and its cellular receptor: A molecular genetic dissection of a virus/receptor affinity interaction

The ability of a virus to attach to a suceptible host cell is of utmost importance for the initiation of viral life cycle. Cell surface proteins called viral receptors mediate the initial steps of virus attachment and uptake. Poliovirus (PV) is one of the most studied animal viruses and its interaction with its cellular receptor, the human poliovirus receptor (hPVR) has been well characterized. This review will present our current understanding of the PV/hPVR interaction at the genetic and biochemical level. In addition, we will also discuss the implicatlions of the PV/hPVR interaction on PV tissue tropism and the evolution of the three PV serotypes.

Authors
Solecki, D; Gromeier, M; Harber, J; Bernhardt, G; Wimmer, E
MLA Citation
Solecki, D, Gromeier, M, Harber, J, Bernhardt, G, and Wimmer, E. "Poliovirus and its cellular receptor: A molecular genetic dissection of a virus/receptor affinity interaction." Journal of Molecular Recognition 11.1-6 (1998): 2-9.
Source
scival
Published In
Journal of Molecular Recognition
Volume
11
Issue
1-6
Publish Date
1998
Start Page
2
End Page
9

Poliovirus and its cellular receptor: a molecular genetic dissection of a virus/receptor affinity interaction.

The ability of a virus to attach to a susceptible host cell is of utmost importance for the initiation of viral life cycle. Cell surface proteins called viral receptors mediate the initial steps of virus attachment and uptake. Poliovirus (PV) is one of the most studied animal viruses and its interaction with its cellular receptor, the human poliovirus receptor (hPVR) has been well characterized. This review will present our current understanding of the PV/hPVR interaction at the genetic and biochemical level. In addition, we will also discuss the implications of the PV/hPVR interaction on PV tissue tropism and the evolution of the three PV serotypes.

Authors
Solecki, D; Gromeier, M; Harber, J; Bernhardt, G; Wimmer, E
MLA Citation
Solecki, D, Gromeier, M, Harber, J, Bernhardt, G, and Wimmer, E. "Poliovirus and its cellular receptor: a molecular genetic dissection of a virus/receptor affinity interaction." J Mol Recognit 11.1-6 (1998): 2-9. (Review)
PMID
10076797
Source
pubmed
Published In
Journal of Molecular Recognition
Volume
11
Issue
1-6
Publish Date
1998
Start Page
2
End Page
9
DOI
10.1002/(SICI)1099-1352(199812)11:1/6<2::AID-JMR380>3.0.CO;2-D

Erratum: A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element (Journal of Virology (1998) 72:7 (5638-5647))

Authors
Das, S; Ott, M; Yamane, A; Tsai, W; Gromeier, M; Lahser, F; Gupta, S; Dasgupta, A
MLA Citation
Das, S, Ott, M, Yamane, A, Tsai, W, Gromeier, M, Lahser, F, Gupta, S, and Dasgupta, A. "Erratum: A small yeast RNA blocks hepatitis C virus internal ribosome entry site (HCV IRES)-mediated translation and inhibits replication of a chimeric poliovirus under translational control of the HCV IRES element (Journal of Virology (1998) 72:7 (5638-5647))." Journal of Virology 72.11 (1998): 9419--.
Source
scival
Published In
Journal of Virology
Volume
72
Issue
11
Publish Date
1998
Start Page
9419-

Genetic recombination of poliovirus in a cell-free system.

Genetic recombination of plus-strand RNA viruses is an important process for promoting genetic variation. By using genetically marked poliovirus RNAs, we have demonstrated that genetic recombination can occur in a cell-free system that generates infective virus from added poliovirus RNA. Recombinant polioviruses were isolated, and the region of crossing over was roughly mapped. Recombinants could be isolated even under conditions where the yield of viruses from one of the parental RNAs was depressed to levels comparable to or less than the yield of recombinant viruses, an observation suggesting that only one of the recombining RNAs needs to be replication-competent. The generation of poliovirus recombinants in a cell-free system offers new possibilities for studying recombination and evolution of RNA viruses.

Authors
Duggal, R; Cuconati, A; Gromeier, M; Wimmer, E
MLA Citation
Duggal, R, Cuconati, A, Gromeier, M, and Wimmer, E. "Genetic recombination of poliovirus in a cell-free system." Proc Natl Acad Sci U S A 94.25 (December 9, 1997): 13786-13791.
PMID
9391105
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
94
Issue
25
Publish Date
1997
Start Page
13786
End Page
13791

Determinants of poliovirus neurovirulence.

Authors
Gromeier, M; Mueller, S; Solecki, D; Bossert, B; Bernhardt, G; Wimmer, E
MLA Citation
Gromeier, M, Mueller, S, Solecki, D, Bossert, B, Bernhardt, G, and Wimmer, E. "Determinants of poliovirus neurovirulence." J Neurovirol 3 Suppl 1 (May 1997): S35-S38.
PMID
9179789
Source
pubmed
Published In
Journal of NeuroVirology
Volume
3 Suppl 1
Publish Date
1997
Start Page
S35
End Page
S38

Internal ribosomal entry site substitution eliminates neurovirulence in intergeneric poliovirus recombinants.

Neuropathogenicity of poliovirus can be attenuated by mutations in the internal ribosomal entry site (IRES) within the 5' nontranslated region of its genome. The Sabin vaccine strains used in prevention of poliomyelitis carry such mutations in their IRES elements. In addition, mutations within the structural and nonstructural proteins of Sabin strains may equally contribute to the attenuation phenotype. Despite their effectiveness as vaccines, the Sabin strains retain a neuropathogenic potential in animal models for poliomyelitis and, at a very low rate, they can cause poliomyelitis in vaccine recipients. The elimination of the neurocytopathic phenotype was achieved through the exchange of the entire poliovirus IRES with its counterpart from human rhinovirus type 2 without affecting growth properties in nonneuronal cells. The attenuating effect of the human rhinovirus type 2 IRES within the context of a poliovirus genome has been mapped to the 3' portion of this genetic element.

Authors
Gromeier, M; Alexander, L; Wimmer, E
MLA Citation
Gromeier, M, Alexander, L, and Wimmer, E. "Internal ribosomal entry site substitution eliminates neurovirulence in intergeneric poliovirus recombinants." Proc Natl Acad Sci U S A 93.6 (March 19, 1996): 2370-2375.
PMID
8637880
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
93
Issue
6
Publish Date
1996
Start Page
2370
End Page
2375

The human poliovirus receptor. Receptor-virus interaction and parameters of disease specificity.

The host range of poliovirus is determined by the expression of the hPVR, a member of the immunoglobulin superfamily. We characterized hPVR proteins biochemically and found them to be complex-type glycoproteins. The outermost V-like domain of three extracellular domains harbors the PVR function. A panel of single or multiple amino acid exchanges were introduced throughout this domain in order to localize regions involved in virus-receptor interactions. Putative contact amino acids were found to reside in the C'C"D and DE regions. Binding and uptake of poliovirus paralleled virus replication in all mutants tested suggesting that virus binding was affected without abrogating the ability to mediate subsequent events in the infection. Although the primate PVR is essential in conferring susceptibility to poliovirus infection, certain strains can induce neurological disease in rodents. Mouse neurovirulent PV isolates of divergent serotypical origin each provoked a distinctive, characteristic neurological syndrome upon intracerebral infection of wild-type mice. We analyzed clinical and histopathological features of diffuse encephalomyelitis caused by these PV strains and compared the condition with poliomyelitis in mice transgenic for the hPVR. Diffuse PV encephalomyelitis in wild-type mice could be distinguished clinically and histopathologically from hPVR-mediated poliomyelitis in trangenic mice. We localized the determinants of mouse neurovirulence of PV1(LS-a), a derivative of PV1 (Mahoney), in a portion of the viral genome encompassing parts of the capsid protein VP1 as well as the nonstructural protein 2A. Mouse neuropathogenicity could possibly be conferred by reduced particle stability of PV1(LS-a) inasmuch as we found particles to be thermolabile.

Authors
Gromeier, M; Lu, HH; Bernhardt, G; Harber, JJ; Bibb, JA; Wimmer, E
MLA Citation
Gromeier, M, Lu, HH, Bernhardt, G, Harber, JJ, Bibb, JA, and Wimmer, E. "The human poliovirus receptor. Receptor-virus interaction and parameters of disease specificity." Ann N Y Acad Sci 753 (May 25, 1995): 19-36.
PMID
7611627
Source
pubmed
Published In
Annals of the New York Academy of Sciences
Volume
753
Publish Date
1995
Start Page
19
End Page
36

Mouse neuropathogenic poliovirus strains cause damage in the central nervous system distinct from poliomyelitis.

Poliomyelitis as a consequence of poliovirus infection is observed only in primates. Despite a host range restricted to primates, experimental infection of rodents with certain genetically well defined poliovirus strains produces neurological disease. The outcome of infection of mice with mouse-adapted poliovirus strains has been described previously mainly in terms of paralysis and death, and it was generally assumed that these strains produce the same disease syndromes in normal mice and in mice transgenic for the human poliovirus receptor (hPVR-tg mice). We report a comparison of the clinical course and the histopathological features of neurological disease resulting from intracerebral virus inoculation in normal mice with those of murine poliomyelitis in hPVR-tg mice. The consistent pattern of clinical deficits in poliomyelitic transgenic mice contrasted with highly variable neurologic disease that developed in mice infected with different mouse-adapted polioviruses. Histopathological analysis showed a diffuse encephalomyelitis induced by specific poliovirus serotype 2 isolates in normal mice, that affected neuronal cell populations without discrimination, whereas in hPVR-tg animals, damage was restricted to spinal motor neurons. Mouse neurovirulent strains of poliovirus type 2 differed from mouse neurovirulent poliovirus type 1 derivatives in their ability to induce CNS lesions. Our findings indicate that the characteristic clinical appearance and highly specific histopathological features of poliomyelitis are mediated by the hPVR.(ABSTRACT TRUNCATED AT 250 WORDS)

Authors
Gromeier, M; Lu, HH; Wimmer, E
MLA Citation
Gromeier, M, Lu, HH, and Wimmer, E. "Mouse neuropathogenic poliovirus strains cause damage in the central nervous system distinct from poliomyelitis." Microb Pathog 18.4 (April 1995): 253-267.
PMID
7476091
Source
pubmed
Published In
Microbial Pathogenesis
Volume
18
Issue
4
Publish Date
1995
Start Page
253
End Page
267

Dicistronic polioviruses as expression vectors for foreign genes.

We have made use of certain novel genetic elements of picornaviruses termed internal ribosomal entry sites (IRES) to construct a viral RNA with the following genetic order: PV 5' NTR-EMCV IRES-PV ORF-3' NTR (PV, poliovirus; NTR, nontranslated region; EMCV, encephalomyocarditis virus; ORF, open reading frame). Transfection of this RNA into HeLa cells yielded a poliovirus (W1-PNENPO) that contained two heterologous IRES elements (type 1 IRES of PV; type 2 IRES of EMCV) in tandem. The insertion of foreign coding sequences into the genome of W1-PNENPO between the IRES elements yielded viable polioviruses with the gene order PV 5' NTR-foreign ORF-EMCV IRES-PV ORF-3' NTR. The foreign ORFs we have employed in this study included the coding region for chloramphenicol acetyltransferase (CAT), or segments of either luciferase or the HIV-1 envelope glycoprotein gp120. W1-PV/V3-3, a dicistronic poliovirus that contained HIV-1-specific sequences that included the V3 domain of gp120, was used to infect transgenic mice (PVR+) that were engineered to express the poliovirus receptor. The genetic stability of the dicistronic viruses and the HIV-1-specific immune response in PVR+ mice after infection with these novel agents are discussed.

Authors
Alexander, L; Lu, HH; Gromeier, M; Wimmer, E
MLA Citation
Alexander, L, Lu, HH, Gromeier, M, and Wimmer, E. "Dicistronic polioviruses as expression vectors for foreign genes." AIDS Res Hum Retroviruses 10 Suppl 2 (1994): S57-S60.
PMID
7865334
Source
pubmed
Published In
AIDS Research and Human Retroviruses
Volume
10 Suppl 2
Publish Date
1994
Start Page
S57
End Page
S60

Kinetics of poliovirus uncoating in HeLa cells in a nonacidic environment.

Lysis of HeLa cells infected with poliovirus revealed intact virus; 135S particles, devoid of VP4 but containing the viral RNA; and 80S empty capsids. During infection the kinetics of poliovirus uncoating showed a continuous decrease of intact virus, while the number of 135S particles and empty shells increased. After 1.5 h of infection conformational transition to altered particles resulted in complete disappearance of intact virions. To investigate the mechanism of poliovirus uncoating, which has been suggested to depend on low pH in endosomal compartments of cells, we used lysosomotropic amines to raise the pH in these vesicles. In the presence of ammonium chloride, however, the kinetics of uncoating were similar to those for untreated cells, whereas in cells treated with methylamine, monensin, or chloroquine, uncoating was merely delayed by about 30 min. This effect could be attributed to a delay of virus entry into cells after treatment with methylamine and monensin, whereas chloroquine stabilized the viral capsid itself. Thus, elevation of endosomal pH did not affect virus uncoating. We therefore propose a mechanism of poliovirus uncoating which is independent of low pH.

Authors
Gromeier, M; Wetz, K
MLA Citation
Gromeier, M, and Wetz, K. "Kinetics of poliovirus uncoating in HeLa cells in a nonacidic environment." J Virol 64.8 (August 1990): 3590-3597.
PMID
2164587
Source
pubmed
Published In
Journal of virology
Volume
64
Issue
8
Publish Date
1990
Start Page
3590
End Page
3597
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